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<title>bioRxiv Subject Collection: Biochemistry</title>
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This feed contains articles for bioRxiv Subject Collection "Biochemistry"
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<link>https://www.biorxiv.org</link>
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<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.15.718671v1?rss=1">
<title>
<![CDATA[
ARMH3 acts as a central scaffold at the Golgi/TGN through interactions with Arl5, GBF1, and PI4KB 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.15.718671v1?rss=1
</link>
<description><![CDATA[
The armadillo repeat protein ARMH3 regulates the activity and localisation of the Golgi resident lipid kinase phosphatidylinositol 4 kinase III{beta} (PI4KB) and the Golgi-specific brefeldin A-resistance guanine nucleotide exchange factor 1 (GBF1) that activates Arf1. ARMH3 localises to the trans Golgi network (TGN) via the GTPase Arl5. We used hydrogen deuterium exchange mass spectrometry (HDX-MS) and AI-enabled modeling to define the interfaces of ARMH3 with its binding partners Arl5, PI4KB, and GBF1. The ARMH3-Arl5 interface was determined to consist of the N and C termini of ARMH3, with Arl5 binding causing conformational changes in ARMH3 located at a shared PI4KB/GBF1 interface. Both PI4KB and GBF1 form mutually exclusive complexes with ARMH3, with GBF1 binding to ARMH3 through a disordered loop we have named the ARMH3 binding region (ABR). The ARMH3 interfaces in PI4KB and GBF1 contain phosphosites, with the phosphomimetic mutation of GBF1 blocking complex formation. These findings provide new insights into the role of ARMH3 as a master coordinator of GTPase and phosphoinositide signaling at the Golgi/TGN.
]]></description>
<dc:creator><![CDATA[ Scott, M. K., Klynsoon, G. C., Walsh, E. E., Suresh, S., Nyvall, H. G., Burke, J. E. ]]></dc:creator>
<dc:date>2026-04-17</dc:date>
<dc:identifier>doi:10.64898/2026.04.15.718671</dc:identifier>
<dc:title><![CDATA[ARMH3 acts as a central scaffold at the Golgi/TGN through interactions with Arl5, GBF1, and PI4KB]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.15.717356v1?rss=1">
<title>
<![CDATA[
Native entanglement misfolding contributes to age-associated structural changes across the Saccharomyces cerevisiae proteome 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.15.717356v1?rss=1
</link>
<description><![CDATA[
Aging at the subcellular level involves the simultaneous decline in the cell's ability to maintain protein homeostasis and rise in misfolded proteins through a positive feedback loop. Here, we test if a widespread class of protein misfolding could contribute to proteome aging by examining if statistical associations exist between age-related changes in protein structure, measured by limited proteolysis mass spectrometry data of the aging Saccharomyces cerevisiae proteome, with structural annotations and molecular simulations. We find that globular proteins that are likely to exhibit entanglement misfolding are 121% more likely to exhibit age-related structural changes, and these changes are 59% more likely to be localized to natively entangled regions. Proteins containing native entanglements are seven-fold more likely to misfold, according to simulations, and populate long-lived, near-native misfolded states. Thus, the age-related structural changes in yeast proteins can be explained in part by the accumulation of misfolded proteins involving entanglements.
]]></description>
<dc:creator><![CDATA[ Vu, Q. V., Sitarik, I., Nissley, D. A., O'Brien, E. P. ]]></dc:creator>
<dc:date>2026-04-17</dc:date>
<dc:identifier>doi:10.64898/2026.04.15.717356</dc:identifier>
<dc:title><![CDATA[Native entanglement misfolding contributes to age-associated structural changes across the Saccharomyces cerevisiae proteome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.16.718858v1?rss=1">
<title>
<![CDATA[
The Effects of Phosphorylation on the Structure and Function of Motif A, an Intrinsically Disordered Region within SIRT1 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.16.718858v1?rss=1
</link>
<description><![CDATA[
The NAD+ dependent deacetylase sirtuin-1 (SIRT1) is known to elicit cellular defenses against aging, cancer, and other aberrant pathologies. Previous studies have identified an intrinsically disordered region of SIRT1 comprised of N-terminal residues 1-52, herein referred to as motif A, which activates SIRT1 activity, likely through intramolecular interactions. Additionally, phosphorylation of N-terminal residues Ser27 and Ser47 has been shown to be important for regulating SIRT1 activity and stability. The lack of in vitro characterization of these effects hampers our further understanding of the role of motif A in SIRT1 regulation. In this study, we elucidate the role phosphorylation plays in motif A structure as well as its regulatory effects on SIRT1 activity against Ac-p65. We find that phosphomimetic mutation at Ser27 significantly increases the activation effect of motif A towards SIRT1. This result is supported by molecular dynamics simulations of the phosphomimetics, which reveal stabilization of different transient structures for motif A depending on whether Ser27 and Ser47 have been modified. A key finding suggested by this study is that phosphorylation of S27 appears to activate SIRT1 by causing motif A, which is intrinsically disordered in the WT, to fold into an ordered structure. This conclusion is based on both the experimental findings and simulation results. These findings contribute to our understanding of SIRT1 regulation, specifically the role played by phosphorylation within the N-terminal disordered region.
]]></description>
<dc:creator><![CDATA[ Richter, S. M., Bui, H.-L., Chen, A., Tannous, C., Butler, B. R., Bennett, S. D., Nguyen, S. Q.-a., Prado, J., Mohamed, A., DuBois, I. A., Tadros, E., Thai, N. T., Lima Guan, S., Peralta, C. M., Kwong, A., Hawk, L. M. L., Grazioli, G., Wang, N. ]]></dc:creator>
<dc:date>2026-04-17</dc:date>
<dc:identifier>doi:10.64898/2026.04.16.718858</dc:identifier>
<dc:title><![CDATA[The Effects of Phosphorylation on the Structure and Function of Motif A, an Intrinsically Disordered Region within SIRT1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.14.718580v1?rss=1">
<title>
<![CDATA[
Integrated bioinformatics and single-cell analysis identifies vascular aging-related hub genes and immune drivers in atherosclerosis. 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.14.718580v1?rss=1
</link>
<description><![CDATA[
Atherosclerosis (AS) is a chronic inflammatory disease closely linked to vascular senescence, yet the specific molecular mechanisms connecting aging processes to AS pathogenesis remain incompletely understood. This study integrated transcriptomic data from GEO datasets (GSE100927 and GSE43292) to identify vascular aging-related differentially expressed genes (VARDEGs). Following batch effect correction, 28 VARDEGs were screened and subjected to functional enrichment, protein-protein interaction (PPI) network analysis, and immune infiltration assessment. Seven hub genes (MMP9, APOE, TNF, ICAM1, PPARG, CYBA, and NCF2) were identified and experimentally validated via qRT-PCR, confirming their significant upregulation in AS samples. Receiver operating characteristic (ROC) analysis demonstrated high diagnostic accuracy for six of these genes (AUC > 0.7), with TNF exhibiting superior performance. Immune infiltration analysis revealed profound alterations in 28 immune cell types, particularly monocytes and T cells, which correlated strongly with hub gene expression. Furthermore, single-cell RNA sequencing analysis (GSE184073) localized the expression of core genes predominantly to monocytes and T cells, highlighting TNF overexpression in T cells as a potential critical driver. Finally, molecular docking simulations suggested that curcumin exhibits strong binding affinity to these hub genes, particularly PPARG, providing a mechanistic basis for its therapeutic potential. Collectively, this study elucidates the landscape of vascular aging-related genes in AS, identifies novel diagnostic biomarkers, and proposes potential therapeutic targets involving immune modulation and natural compounds.
]]></description>
<dc:creator><![CDATA[ Wu, J., Chen, X., Zhou, K., Wang, W. ]]></dc:creator>
<dc:date>2026-04-17</dc:date>
<dc:identifier>doi:10.64898/2026.04.14.718580</dc:identifier>
<dc:title><![CDATA[Integrated bioinformatics and single-cell analysis identifies vascular aging-related hub genes and immune drivers in atherosclerosis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.15.718621v1?rss=1">
<title>
<![CDATA[
A new stress-response pathway in Mycobacterium tuberculosis 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.15.718621v1?rss=1
</link>
<description><![CDATA[
Metabolic adaptations are key in the virulence of the pathogen Mycobacterium tuberculosis (Mtb). However, our current understanding of these adaptations is limited to common pathways in central carbon metabolism. Here we apply an untargeted bottom-up approach to discover unknown stress-responsive metabolites and their biosynthetic enzyme. We show that upon exposure to hypoxia, nitric oxide and activated macrophages, Mtb rapidly produces high levels of an unknown metabolite that we identify as 6'-{gamma}-amino-butyric acid-trehalose (GABA-trehalose). Formation of millimolar GABA-trehalose levels under these stresses is driven by a rapid rise in GABA, which is also excreted. We demonstrate that GABA-trehalose is produced from GABA and trehalose by the uncharacterized ATP-grasp enzyme Rv1722, involving a carboxylate-hydroxyl ligation that is non-canonical for ATP-grasp enzymes. Phylogenetic analyses demonstrate that the gene rv1722 is present in most slow-growing mycobacteria but absent in most rapid-growing mycobacteria. While the role of GABA-trehalose in Mtb metabolism remains unclear, we postulate that the increased NADH/NAD+ ratio under hypoxia and nitric oxide exposure promotes GABA formation and inhibits its breakdown, leading to GABA accumulation and excretion. Rv1722-driven coupling of GABA and trehalose constitutes an alternative to excretion that conserves carbon and nitrogen. Taken together, our bottom-up approach reveals a new stress-response pathway in Mtb that rapidly produces large quantities of GABA-trehalose. These findings extend our knowledge of the metabolic adaptations that a major human pathogen utilizes in response to immune system-imposed stresses.
]]></description>
<dc:creator><![CDATA[ van der Velden, P. M., Merx, J., van Dijk, L., Roos, F., Brake, J., Berben, T., White, P. B., de Graaf, R. M., Terschlusen, E., van Weerdenburg, I. J., Zhang, B. H., van Crevel, R., van Niftrik, L., van Ingen, J., Boltje, T. J., Jansen, R. S. ]]></dc:creator>
<dc:date>2026-04-16</dc:date>
<dc:identifier>doi:10.64898/2026.04.15.718621</dc:identifier>
<dc:title><![CDATA[A new stress-response pathway in Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.15.718664v1?rss=1">
<title>
<![CDATA[
Acyl-enzyme dynamics, tautomerisation and hydration regulate turnover of carbapenem antibiotics by the OXA-48 β-lactamase 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.15.718664v1?rss=1
</link>
<description><![CDATA[
OXA-48 is a globally disseminated class D serine {beta}-lactamase that efficiently confers resistance to a range of {beta}-lactam antibiotics, including carbapenems, the most potent such agents versus Enterobacterales (Escherichia coli and relatives). Here we characterise the interactions of OXA-48 with the acyl-enzyme complex intermediates formed on its reaction with the carbapenems meropenem and ertapenem using X-ray crystallography and molecular dynamics (MD) simulations. X-ray crystal structures identify acyl-enzymes in both the {Delta}1-imine and {Delta}2-enamine pyrroline tautomeric forms. MD simulations show the epimeric {Delta}2 tautomers of meropenem and ertapenem to more frequently adopt binding poses competent for hydrolysis, i.e. with an appropriate orientation of the carbapenem 6-hydroxyethyl group and positioning of the water molecule required for deacylation; the results indicate that the {Delta}2 tautomers are preferred for deacylation over the {Delta}1-tautomer. MD simulations based on the crystal structures show that, compared to OXA-48, acyl-enzyme complexes of OXA-519 (a natural OXA-48 variant with a single Val120Leu substitution adjacent to the catalytic general base) more frequently sampled conformations favouring hydrolysis, or formation of the alternative {beta}-lactone deacylation product. MD simulations of complexes derived from quantum mechanics/molecular mechanics (QM/MM) simulations show the meropenem-derived {beta}-lactone product is better retained in the OXA-48 active site than hydrolysed meropenem, consistent with reversible {beta}-lactone formation. Overall, our results demonstrate how acyl-enzyme tautomerisation, dynamics and hydration collectively modulate degradation of 1{beta}-methyl carbapenems by class D {beta}-lactamases of the OXA-48 group, and how subtle changes in active site structure potentiate such effects in the OXA-519 variant.
]]></description>
<dc:creator><![CDATA[ Hoff, J. F., Beer, M., Hinchliffe, P., Tooke, C. L., Schofield, C. J., van der Kamp, M. W., Mulholland, A. J., Spencer, J. ]]></dc:creator>
<dc:date>2026-04-16</dc:date>
<dc:identifier>doi:10.64898/2026.04.15.718664</dc:identifier>
<dc:title><![CDATA[Acyl-enzyme dynamics, tautomerisation and hydration regulate turnover of carbapenem antibiotics by the OXA-48 β-lactamase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.14.718489v1?rss=1">
<title>
<![CDATA[
Cephalopod Genome Expansion Drives Broader Reflectin Domain Boundaries 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.14.718489v1?rss=1
</link>
<description><![CDATA[
As more genome libraries become accessible across multiple cephalopod species, long standing assumptions about the molecular basis of their dynamic optical systems are being revisited. For example, cephalopod-specific reflectin proteins now appear far more variable as newly annotated sequences diverge from the canonical features that have long defined this protein family. In this perspective, we discuss these nuances and introduce a theoretical framework that expands the reflectin domain classification while preserving specificity across 141 known and previously uncharacterized reflectin sequences from ten cephalopod species. By combining this broadened domain architecture with quantitative, bioinformatics driven metrics, we establish a generalizable framework that reliably identifies reflectins across their full sequence diversity, allowing for deeper structural comparisons and future functional discovery.
]]></description>
<dc:creator><![CDATA[ Armendarez, O. J., Bower, D. Q., Flynn, K. R., Bergman, M. R., Albertin, C. B., Deravi, L. F. ]]></dc:creator>
<dc:date>2026-04-16</dc:date>
<dc:identifier>doi:10.64898/2026.04.14.718489</dc:identifier>
<dc:title><![CDATA[Cephalopod Genome Expansion Drives Broader Reflectin Domain Boundaries]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.15.718697v1?rss=1">
<title>
<![CDATA[
Structural basis of chaperone mechanisms in cells and the evolutionary emergence of the protein world 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.15.718697v1?rss=1
</link>
<description><![CDATA[
How chaperones mediate protein folding in the crowded cell environment remains poorly understood. To gain insight, we developed CHAP-SEQ to examine how chaperones affect protein folding in cells at high throughput and amino acid resolution. Performing CHAP-SEQ using three chaperone proteins and one chaperone RNA reveals distinct modes of folding assistance. Chaperone proteins act preferentially on hydrophobic core residues, whereas chaperone RNA primarily targets structural or dynamic signatures. Furthermore, while the chaperone RNA has little preference for clients' baseline foldability, the chaperone proteins favor clients with greater intrinsic foldability. These differences are consistent with an evolutionary hypothesis in which greater chaperone complexity played a role in the formation of stable hydrophobic cores, suggesting a potential link between chaperone function and the evolution of protein folding.
]]></description>
<dc:creator><![CDATA[ Son, A., Durso, C., Whitehead, T. A., Horowitz, S. ]]></dc:creator>
<dc:date>2026-04-16</dc:date>
<dc:identifier>doi:10.64898/2026.04.15.718697</dc:identifier>
<dc:title><![CDATA[Structural basis of chaperone mechanisms in cells and the evolutionary emergence of the protein world]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.13.718317v1?rss=1">
<title>
<![CDATA[
Cooperative Control of Arrestin Activation By Membrane Lipids And Phosphorylation Barcodes 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.13.718317v1?rss=1
</link>
<description><![CDATA[
Arrestins regulate G protein-coupled receptor (GPCR) signaling by binding phosphorylated receptors embedded in lipid bilayers, yet how receptor phosphorylation and membrane composition cooperate to control arrestin activation remains unclear. Here, we reconstitute this interplay using N-terminally palmitoylated phosphopeptides tethered to nanodiscs of defined lipid composition and quantitatively measure arrestin-2 (Arr2) activation and membrane engagement. We find that both receptor phosphorylation and the lipid environment are essential for robust Arr2 activation, with phosphoinositides (PIPs) and other anionic lipids facilitating Arr2 activation and membrane association through distinct mechanisms. Systematic profiling of phosphorylation barcodes derived from atypical chemokine receptor 3 (ACKR3) and vasopressin receptor 2 (V2R) identifies phospho-motifs that potently activate Arr2. Moreover, the position of these motifs relative to the membrane determines Arr2 engagement, supporting a model of regional phosphorylation barcodes. Genome-wide motif analysis further links the phosphorylation barcode to predicted arrestin coupling strength and classification into Class A or Class B GPCRs. Finally, lipidated phosphopeptides inhibit GPCR-Arr2 interactions in live cells and enable structural characterization of Arr2-phosphopeptide complexes by cryo-electron microscopy, establishing a membrane-integrated framework for decoding arrestin response.
]]></description>
<dc:creator><![CDATA[ Aydin, Y., Zhuo, Y., Yen, Y.-C., Chen, C.-L., Klug, C. S., Marchese, A., Chen, Q. ]]></dc:creator>
<dc:date>2026-04-16</dc:date>
<dc:identifier>doi:10.64898/2026.04.13.718317</dc:identifier>
<dc:title><![CDATA[Cooperative Control of Arrestin Activation By Membrane Lipids And Phosphorylation Barcodes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.13.718141v1?rss=1">
<title>
<![CDATA[
Photoacoustic Fingerprinting for Robust Molecular Imaging 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.13.718141v1?rss=1
</link>
<description><![CDATA[
Quantitative molecular imaging in photoacoustics is fundamentally limited by the ill-posed nature of spectral unmixing, where spectral overlap, noise, and unknown fluence introduce bias in conventional inversion-based methods. We introduce photoacoustic fingerprinting (PAF), a framework that reframes spectral unmixing as a fingerprint recognition problem. PAF interprets multispectral signals as high-dimensional fingerprints encoding both molecular composition and measurement distortions. Inspired by magnetic resonance fingerprinting, PAF uses a recurrent neural network trained on synthetic data spanning realistic mixtures, noise levels, and fluence variations to directly infer molecular concentrations from spectral shape. PAF enables accurate and robust quantification in regimes where conventional methods break down, including low signal-to-noise conditions, spectrally correlated mixtures, and unknown fluence distortions. In controlled simulations, PAF consistently outperformed non-negative least squares, with the largest gains observed for spectrally overlapping chromophores such as collagen. In phantom studies, PAF improved molecular specificity by correctly localizing collagen and recovering water contrast despite similar spectral reconstructions. In ex vivo mouse livers, PAF detected lipid accumulation associated with steatosis, and in human arteries, it identified molecular signatures consistent with thrombus and lipid-rich plaque. These results establish PAF as a generalizable framework for label-free molecular imaging and a promising step toward quantitative photoacoustic diagnostics.
]]></description>
<dc:creator><![CDATA[ McGarraugh, C., Menozzi, L., Yao, R., Eng-Wu, D., Nguyen, V. T., Cho, S.-W., Francis, S., Yao, J. ]]></dc:creator>
<dc:date>2026-04-15</dc:date>
<dc:identifier>doi:10.64898/2026.04.13.718141</dc:identifier>
<dc:title><![CDATA[Photoacoustic Fingerprinting for Robust Molecular Imaging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.14.718415v1?rss=1">
<title>
<![CDATA[
Molecular Architecture of Cryptococcus Cell Walls Reveals Species-Specific Chitosan-Dependent Remodeling 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.14.718415v1?rss=1
</link>
<description><![CDATA[
Cryptococcus neoformans and Cryptococcus gattii are fungal pathogens that cause life-threatening infections, including cryptococcal meningitis. A distinctive feature of the cryptococcal cell wall is the extensive deacetylation of chitin to chitosan, a modification that is essential for virulence but whose structural role in cell-wall organization remains poorly understood. Here, we analyzed the cell walls of wild-type strains of both species and their avirulent chitosan-deficient mutants, which serve as vaccine candidates. Loss of chitosan disrupted cell morphology and altered cell-wall ultrastructure, with more pronounced defects in C. neoformans. Solid-state NMR revealed that aggregated -1,3-glucans form the principal rigid domain of the cell wall in both species and are closely associated with chitin microfibrils, whereas surrounding {beta}-glucans and mannoproteins constitute a more dynamic matrix. Chitosan modulates hydration and flexibility, and its loss increases chitin exposure and triggers species-specific remodeling of the polysaccharide network. In C. neoformans, chitosan depletion increased -1,3-glucan content and reduced {beta}-glucan levels, whereas C. gattii selectively lost one -1,3-glucan subtype while maintaining {beta}-glucan levels. Although capsule production remained intact, chitosan deficiency altered glucuronoxylomannan linkage patterns and mannoprotein composition. These findings reveal how chitosan organizes cryptococcal cell-wall architecture and highlight distinct structural adaptation strategies among pathogenic Cryptococcus species.
]]></description>
<dc:creator><![CDATA[ Ankur, A., Upadhya, R., Doosti, M., Ferreira, D., Xie, L., Hung, I., Lodge, J., Wang, T. ]]></dc:creator>
<dc:date>2026-04-15</dc:date>
<dc:identifier>doi:10.64898/2026.04.14.718415</dc:identifier>
<dc:title><![CDATA[Molecular Architecture of Cryptococcus Cell Walls Reveals Species-Specific Chitosan-Dependent Remodeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.13.718024v1?rss=1">
<title>
<![CDATA[
ChemCell: Chemical Tethering of Large Biomolecules to Cell Surfaces through Diels-Alder Ligation 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.13.718024v1?rss=1
</link>
<description><![CDATA[
Chemical engineering of cellular surfaces offers a powerful tool to endow cells with new properties and functions, yet methods for adding complex biomolecules to cell membranes remain underdeveloped. While technologies for targeted degradation of cell surface components are advancing, few approaches exist that allow efficient attachment of large biomolecules. To address this, we introduce ChemCell technology--a platform for non-genetic surface engineering of cells via metabolic installation of trans-cyclooctene (TCO) groups. These TCO groups undergo rapid and selective reactions with tetrazine-modified biomolecules, enabling the efficient tethering of diverse functional groups. This method allows for the precise attachment of proteins, peptides, enzymes, oligonucleotides, therapeutic antibodies, and large protein complexes to cell surfaces, expanding the toolbox for cellular modification and offering new possibilities for cell-based therapies and diagnostics.
]]></description>
<dc:creator><![CDATA[ Dzijak, R., Bellova, S., Kovalova, A., Slachtova, V., Rahm, M., Berankova, A., Pohl, R., Vrabel, M. ]]></dc:creator>
<dc:date>2026-04-15</dc:date>
<dc:identifier>doi:10.64898/2026.04.13.718024</dc:identifier>
<dc:title><![CDATA[ChemCell: Chemical Tethering of Large Biomolecules to Cell Surfaces through Diels-Alder Ligation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.13.718121v1?rss=1">
<title>
<![CDATA[
A miniaturized MR1 metabolite display system with native-like protein features 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.13.718121v1?rss=1
</link>
<description><![CDATA[
Major histocompatibility complex class I-related protein 1 (MR1) presents metabolite-derived antigens to mucosal-associated invariant T (MAIT) cells and other MR1-restricted T cells, playing a critical role in immune surveillance during infection and disease. Biochemical and structural studies of MR1 have been limited by the intrinsic instability of the molecule, which requires both ligand binding and association with beta-2-microglobulin ({beta}2m) for proper folding and stability. Here, we adapt MR1 to the SMART protein platform to generate a minimalistic system for studying MR1 ligand presentation and T cell receptor (TCR) recognition. SMART-MR1 consists of the MR1 1/2 ligand-binding platform fused to a helical stabilizing domain that functionally replaces the 3 and {beta}2m domains, resulting in a truncated protein that preserves the architecture of the antigen-binding groove. We show that SMART-MR1 can be efficiently produced recombinantly and retains the ability to bind chemically diverse classes of MR1 ligands. The reduced size of SMART-MR1 enables amide-based solution NMR experiments, and its simplified structure allows for ligand screening using fluorescence polarization. Importantly, SMART-MR1 maintains binding to the MAIT-derived A-F7 TCR, as confirmed by isothermal titration calorimetry. Finally, cryo-EM structural analysis of SMART-MR1/5-OP-RU bound to A-F7 revealed that ligand presentation and TCR recognition are nearly identical to those observed in native MR1. Together, these results establish SMART-MR1 as a minimal yet native-like system, expanding the experimental toolkit available for studying MR1 interactions and facilitating future efforts aimed at targeting MR1 pathways.

Significance StatementMR1 is a highly conserved antigen-presenting molecule that enables T cells to detect metabolite signals from microbial infections and host metabolism. Despite its importance in immunity, mechanistic studies of MR1 have been limited by the instability of the native protein. We developed a simplified and stabilized version of MR1 that preserves the ligand-binding platform while eliminating structural elements that complicate biochemical analysis. This minimal system retains native-like antigen presentation and T cell receptor (TCR) recognition, while enabling experimental approaches that are difficult with full-length MR1. By lowering technical barriers to studying MR1-ligand and MR1-TCR interactions, this platform provides a versatile tool for exploring how antigens shape immune responses and for accelerating discovery of therapeutic strategies targeting MR1.
]]></description>
<dc:creator><![CDATA[ Rotsides, P., Shinde, O., Danon, J. N., Sgourakis, N. ]]></dc:creator>
<dc:date>2026-04-15</dc:date>
<dc:identifier>doi:10.64898/2026.04.13.718121</dc:identifier>
<dc:title><![CDATA[A miniaturized MR1 metabolite display system with native-like protein features]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.12.717948v1?rss=1">
<title>
<![CDATA[
Dietary Oxysterols Reprogram Hepatic Lipid Metabolism and Reshape the Gut Metabolome-Microbiome Interface 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.12.717948v1?rss=1
</link>
<description><![CDATA[
Dietary oxysterols are biologically active cholesterol oxidation products ubiquitous in Western diets, yet their systemic effects on host metabolism and the gut microbiome remain largely unexplored. Here, we employed an integrated multi-omics approach - shotgun metagenomics, quantitative proteomics, untargeted metabolomics, and bulk RNA-seq - to characterize the impact of DOxS exposure on the gut-liver axis in rats fed a Western diet (WD vs. WD-DOxS). Hepatic proteomics revealed near-complete suppression of the mevalonate/cholesterol biosynthesis pathway, particularly in males, while de novo lipogenesis enzymes (Scd1, Fasn, Plin2) were paradoxically upregulated, consistent with dual oxysterol signaling through SREBP inhibition and LXR activation. Bile acid synthesis was concurrently suppressed, confirmed by metabolomics. Strikingly, RNA-seq across liver, heart, and brain detected virtually no differentially expressed genes, establishing that DOxS act predominantly through post-transcriptional mechanisms. In the gut, DOxS increased microbial -diversity while depleting Limosilactobacillus reuteri, with concomitant loss of the barrier-protective metabolite 3-indoleacrylic acid. Tissue-specific responses were widespread, with liver and colon frequently mounting opposing metabolic and immune responses to the same dietary challenge. Cross-omics integration revealed convergent microbiome-metabolite axes connecting microbial remodeling to both hepatic lipid reprogramming and colonic barrier disruption. These findings reposition dietary oxysterols from food-quality markers to active modulators of the gut-liver axis, with implications for metabolic disease and intestinal barrier integrity.
]]></description>
<dc:creator><![CDATA[ Maldonado Pereira, L., Mutawi, T. M. A., Singh, A., Sanderson, B., Rekowski, M. J., Barnaba, C., Medina Meza, I. G. ]]></dc:creator>
<dc:date>2026-04-14</dc:date>
<dc:identifier>doi:10.64898/2026.04.12.717948</dc:identifier>
<dc:title><![CDATA[Dietary Oxysterols Reprogram Hepatic Lipid Metabolism and Reshape the Gut Metabolome-Microbiome Interface]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.13.718289v1?rss=1">
<title>
<![CDATA[
Small Molecule Regulation of CLOCK:BMAL1 DNA Binding Activity 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.13.718289v1?rss=1
</link>
<description><![CDATA[
CLOCK:BMAL1 is a bHLH-PAS transcription factor complex that utilizes its bHLH (basic helix-loop-helix) domains to bind E-box motifs in DNA and tandem PAS (PER-ARNT-SIM) domains to heterodimerize and interact with regulatory proteins to generate circadian rhythms. PAS domains are evolutionarily conserved modules that frequently bind small molecule ligands within buried cavities to perform sensory and signal transduction functions. CLOCK and BMAL1 PAS domains have cavities that could be leveraged to regulate the transcription factor, and consequently, the circadian clock. Using NMR spectroscopy, we identified small molecules that bind within a cavity inside the PAS-A domain of CLOCK and its paralog NPAS2, which sits at an important flexible junction in the structured core of the heterodimer. We identified a gatekeeping mutant in the core of CLOCK PAS-A that significantly decreased ligand binding affinity. High-pressure NMR studies showed that ligand binding or the gatekeeping mutant significantly stabilized the domain. Finally, we showed that ligands induced dose-dependent displacement of CLOCK:BMAL1 from DNA in vitro. Together, these data demonstrate that small molecules can regulate DNA binding by the circadian transcription factor CLOCK:BMAL1 through occupancy of a PAS domain cavity.
]]></description>
<dc:creator><![CDATA[ Sharma, D., Boral, S., West, E., Kressman, M., Franco, I., Amezcua, C. A., Tripathi, S., Lee, H.-W., Favaro, D. C., Gardner, K. H., Partch, C. L. ]]></dc:creator>
<dc:date>2026-04-14</dc:date>
<dc:identifier>doi:10.64898/2026.04.13.718289</dc:identifier>
<dc:title><![CDATA[Small Molecule Regulation of CLOCK:BMAL1 DNA Binding Activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.13.715649v1?rss=1">
<title>
<![CDATA[
Multiscale Physical Effects of CpG Methylation on DNA Mechanics, Nucleosome Wrapping, and Chromatin Condensates 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.13.715649v1?rss=1
</link>
<description><![CDATA[
DNA methylation at CpG dinucleotides is a central epigenetic modification linked to transcriptional repression and heterochromatin formation, yet its physical impact on chromatin organization remains incompletely understood. Here, we integrate single-molecule force spectroscopy, FRAP, and nanorheology to quantify how CpG methylation alters chromatin mechanics across scales. Optical tweezer-based measurements show that CpG methylation modestly increases DNA contour and persistence lengths, indicating a slightly extended and stiffer DNA helix. When assembled into nucleosomes, CpG methylation weakens both outer-and inner turn wrapping, reducing unwrapping forces and per-nucleosome unwrapping energy. On a higher-order assembly scale, although CpG methylated and unmethylated chromatin fibers display similar thresholds for salt-induced phase separation, methylated condensates exhibit markedly reduced internal mobility: FRAP reveals slower chromatin exchange, nanorheology shows increased viscoelasticity and longer relaxation times. Finally, CpG methylation enhances HP1-chromatin interactions, lowering the HP1 concentration required for condensation, despite no change in HP1 binding to naked DNA. Together, these results show that CpG methylation remodels chromatin mechanics by tuning DNA rigidity, nucleosome wrapping energetics, and condensate material properties, providing a multiscale physical framework for how DNA methylation contributes to heterochromatin.
]]></description>
<dc:creator><![CDATA[ Daris, L. C., Rizvi, A., Singh, A. A., Hutchings, J., Lamers, L. A., Montabana, E. A., Kimanius, D., Banerjee, P. R., Eeftens, J. M., Sanulli, S. ]]></dc:creator>
<dc:date>2026-04-14</dc:date>
<dc:identifier>doi:10.64898/2026.04.13.715649</dc:identifier>
<dc:title><![CDATA[Multiscale Physical Effects of CpG Methylation on DNA Mechanics, Nucleosome Wrapping, and Chromatin Condensates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.11.717874v1?rss=1">
<title>
<![CDATA[
Investigating the Consequences of Non-active site Mutations on the Structure, Function and Dynamics of the Molten Globule Enzyme Monomeric Chorismate Mutase 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.11.717874v1?rss=1
</link>
<description><![CDATA[
Intrinsically disordered enzymes serve as useful models to understand their catalytic function against the backdrop of an unstructured protein. The characteristic flexibility in conformation seen in IDPs is a rare occurrence among enzymes and one such enzyme is the engineered protein: monomeric Chorismate Mutase (mCM). mCM surprisingly retains similar enzyme activity as its parent dimeric protein Chorismate Mutase from Methanococcus jannaschii (MjCM) despite losing the ordered globular structure. In this work using a previously demonstrated transition state analogue (TSA), we analyze the structural transitions in mCM during catalysis. Additionally, consequences of three non-active site single point mutations were investigated using CD; Trp-Dansyl FRET measurements using fluorescence lifetime; and time-resolved fluorescence anisotropy measurements; to map the local (near Trp) and global structural transitions in mCM during catalysis. Mutant2 (W24K + C69A); and Mutant3 (W24K + C69A + A6C); revealed a 97 and 89% drop-in activity compared to mCM; quite unlike Mutant1 (W24K, 19% drop). Mutant1 as opposed to Mutant3 was most sensitive to binding of TSA as quantified by structural displacement measured using FRET. This was consistent with an overall globular structure compaction induced by TSA binding in Mutant1 as reflected by a dip in rotational correlation time of Cys-conjugated dansyl probe from 10.3 to 8.4 ns. Our results highlight the critical role of Cys69 residue, that is ~19 [A] away from mCM active site, in influencing the hydrophobic collapse upon substrate binding and subsequent catalytic activity.
]]></description>
<dc:creator><![CDATA[ Biswas, S., Gangadhar, P., Pabbaraja, S., Swaminathan, R. ]]></dc:creator>
<dc:date>2026-04-14</dc:date>
<dc:identifier>doi:10.64898/2026.04.11.717874</dc:identifier>
<dc:title><![CDATA[Investigating the Consequences of Non-active site Mutations on the Structure, Function and Dynamics of the Molten Globule Enzyme Monomeric Chorismate Mutase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.10.717634v1?rss=1">
<title>
<![CDATA[
Coupling of electron-bifurcation modules powers aromatic ring reduction beyond the biological redox window 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.10.717634v1?rss=1
</link>
<description><![CDATA[
Microbial degradation of ubiquitous aromatic compounds is central to the global carbon cycle and bioremediation, yet the intrinsic stability of aromatic rings poses a major barrier to their breakdown. Under strictly anaerobic conditions, class II benzoyl-CoA reductases (BCRII) catalyse the key step of this process, a Birch-like reduction of the aromatic ring to a cyclic diene at a tungsten cofactor. This reaction operates beyond the redox limits of conventional biological electron transfer, yet the mechanism by which BCRII generates such extreme reducing power has remained unclear. Here, high-resolution cryo-electron microscopy, in situ cryo-electron tomography, and enzymatic analyses reveal that the one-MDa BCRII complex from Geobacter metallireducens links two distinct flavin-based electron-bifurcation modules, previously characterised in hydrogenases and heterodisulfide reductases, to drive aromatic ring reduction. Reduced ferredoxin and NADH deliver electrons through sequential confurcation and bifurcation to the catalytic site, while cryo-electron tomography of native cells identifies electron-transferring flavoprotein as the second acceptor directing high-potential electrons into the respiratory chain. These results show that modular FBEB units can be hierarchically assembled to extend metabolic redox capacity, highlighting their versatility as adaptable components for electron transfer pathways.
]]></description>
<dc:creator><![CDATA[ Appel, L., Kumar, A., Tamborrini, D., Pascoa, T. C., Kayastha, K., Ermler, U., Kettler, T., Bohn, S., Reif-Trauttmansdorff, T., Engel, B. D., Schuller, J. M., Boll, M. ]]></dc:creator>
<dc:date>2026-04-13</dc:date>
<dc:identifier>doi:10.64898/2026.04.10.717634</dc:identifier>
<dc:title><![CDATA[Coupling of electron-bifurcation modules powers aromatic ring reduction beyond the biological redox window]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.10.717654v1?rss=1">
<title>
<![CDATA[
Saliva Metabolomics Reveals Distinct Metabolic Signatures in Patients with Chronic Obstructive Pulmonary Disease: A GC-MS-based approach. 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.10.717654v1?rss=1
</link>
<description><![CDATA[
Chronic obstructive pulmonary disease (COPD), a chronic lung disease, involves complex metabolic disturbances that remain poorly characterized using non-invasive matrices. The metabolic alterations associated with cigarette smoke (CS), one of the major drivers of disease progression in COPD patients, have not been explored in detail. This study primarily aimed to investigate the metabolic signatures in COPD patients categorized into smoker (n=15), ex-smoker (n=11), and non-smoker (n=3) subgroups. Utilizing saliva as a noninvasive sample, we identified 26 metabolites with differential expression in smokers and 31 in ex-smokers. However, no such significant alteration was observed in the non-smokers subgroup. The multivariate analysis distinctly separated the COPD subgroups from healthy controls. Additionally, pathway enrichment analysis revealed perturbations in key metabolic pathways, including unsaturated fatty acid biosynthesis, arginine biosynthesis, the tricarboxylic acid (TCA) cycle, and pyruvate metabolism. Moreover, univariate Random forest analysis identified four metabolites (cyclopentanone, tetradecane 4-methyl, acetophenone, and scyllo-inositol) as potential biomarkers distinguishing COPD subgroups from healthy controls. This study offers novel molecular insights into the association of smoking with disease progression and provides a mechanistic understanding of COPD in different subgroups for better management of the disease.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=163 SRC="FIGDIR/small/717654v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator><![CDATA[ Singh, R., Ghosh, S., Yadav, N., Mandal, A. K. ]]></dc:creator>
<dc:date>2026-04-13</dc:date>
<dc:identifier>doi:10.64898/2026.04.10.717654</dc:identifier>
<dc:title><![CDATA[Saliva Metabolomics Reveals Distinct Metabolic Signatures in Patients with Chronic Obstructive Pulmonary Disease: A GC-MS-based approach.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.11.717888v1?rss=1">
<title>
<![CDATA[
Immobilized enzyme-assisted production of recombinant P113 peptide 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.11.717888v1?rss=1
</link>
<description><![CDATA[
Recombinant peptide production was pioneered in the 1970s for the generation of therapeutic peptides, with notable examples including insulin and somatostatin. These early methods required the use of cyanogen bromide (BrCN) for cleavage of the native peptide sequence from a fusion protein. Since that time, while numerous BrCN-dependent peptide methods continue to be reported, the accessibility and cost of site-specific proteases have improved dramatically. These developments have enabled alternative approaches to recombinant peptide generation that obviate the need for BrCN, an environmentally destructive toxin. We recently created an immobilized SUMO protease that can replace BrCN usage in recombinant peptide production workflows by releasing native peptides expressed as part of a SUMO-peptide fusion protein. We have used this approach to generate P113 peptide, the minimal active fragment of the antifungal peptide Histatin 5. In this report, we describe the creation and characterization of this immobilized SUMO protease and its application in the production of experimentally viable quantities of active P113 peptide.
]]></description>
<dc:creator><![CDATA[ Kirkendoll, J. A., Targino Campos, L., Taylor, E. G., Murata, R. M., Hughes, R. M. ]]></dc:creator>
<dc:date>2026-04-13</dc:date>
<dc:identifier>doi:10.64898/2026.04.11.717888</dc:identifier>
<dc:title><![CDATA[Immobilized enzyme-assisted production of recombinant P113 peptide]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.10.717665v1?rss=1">
<title>
<![CDATA[
Detergent-Free Nuclear-Cytoplasmic Fractionation Enables Spatially Resolved PELSA for Enhanced Nuclear Drug Target Identification 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.10.717665v1?rss=1
</link>
<description><![CDATA[
Accurate identification of drug target proteins remain major challenges in proteomics-based target discovery, particularly for low-abundance nuclear proteins that are difficult to detect because of the complexity of whole-cell lysates. Here, we developed a detergent-free nuclear-cytoplasmic fractionation strategy compatible with peptide-centric local stability analysis (PELSA), which markedly improves detection of nuclear drug targets. Using K562 cells, we demonstrated that mild detergent-free fractionation enables high-fidelity nuclear-cytoplasmic separation with minimal cross-contamination. When coupled with PELSA, this workflow significantly increases the number of detected nuclear targets relative to whole-cell analysis. Benchmarking with well-characterized nuclear drugs, including the histone deacetylase inhibitor panobinostat and the RNA polymerase II inhibitor -amanitin, our results showed improved identification of canonical nuclear targets. Broad profiling of staurosporine target further revealed expanded kinase target coverage by combining the results of nuclear and cytoplasmic fraction, with the CLK family kinases detected exclusively in the nuclear fractions. Additionally, we showed that PELSA can also be performed on intact nucleus level. Collectively, these findings establish detergent-free nuclear-cytoplasmic fractionation-PELSA as a robust and scalable strategy for spatially resolved drug target identification, improving sensitivity for nuclear and low-abundance proteins.
]]></description>
<dc:creator><![CDATA[ Cai, D., Zou, K., Wang, J., Zhu, H., Ma, Y., Yang, D., Zhang, X., Yan, J., Zou, L., Wang, K., Ye, M. ]]></dc:creator>
<dc:date>2026-04-13</dc:date>
<dc:identifier>doi:10.64898/2026.04.10.717665</dc:identifier>
<dc:title><![CDATA[Detergent-Free Nuclear-Cytoplasmic Fractionation Enables Spatially Resolved PELSA for Enhanced Nuclear Drug Target Identification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.12.718052v1?rss=1">
<title>
<![CDATA[
Crystal structure and molecular dynamics simulations of rademikibart Fab-IL-4Rα complex reveal biochemical basis for next-generation potent IL-4Rα inhibition in type 2 allergic and inflammatory diseases 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.12.718052v1?rss=1
</link>
<description><![CDATA[
Rademikibart (CBP-201) is a human monoclonal antibody with higher binding affinity to IL-4R compared to dupilumab. Dupilumab is a first-generation interleukin-4 receptor alpha (IL-4R) inhibitor for treating IL-4R-dependent inflammatory disorders, including several dermatologic and respiratory conditions. Rademikibart, however, demonstrated better inhibition of STAT6 intracellular signaling in vitro and similar potency in inhibiting both IL-4 induced TARC release and IL-4 induced B cell activation. To further characterize the molecular function of rademikibart and its differentiation from dupilumab, we determined the crystal structure of the rademikibart fragment antigen binding (Fab) bound to IL-4R at 2.71 [A] resolution and compared this to the 2.82 [A] resolution structure of dupilumab Fab bound to IL-4R. The rotation angle between dupilumab and rademikibart bound to IL-4R is 54.88{degrees}. This rotation enables the binding epitopes of rademikibart, but not dupilumab, on IL-4R to overlap more closely with the conserved binding interface naturally utilized by IL-4 and IL-13 cytokines. Molecular dynamics (MD) studies on rademikibart and dupilumab bound to IL-4R examined the stability of the complexes and effects of amino acid mutations on receptor complex formation. MD simulations demonstrated that the third interface loop (residues 145 to 153 in domain 2) of IL-4R interacts directly with rademikibart, which is absent in the dupilumab/IL-4R complex. This finding is confirmed by increased hydrogen bond interactions at the interface between rademikibart and IL-4R, demonstrating superior binding energy for rademikibart. Through analysis of the x-ray crystallography structures, MD-equilibrated structures, and computational point-mutation analysis of rademikibart, we identified residue Y50 and R55 of the light chain and R97, R99, and Y101 of the heavy chain of rademikibart as key residues interacting with IL-4Rs third interface loop. Our data provides a molecular and structural rationale for the enhanced IL-4R inhibition by rademikibart over dupilumab, confirming rademikibart as an optimized second-generation IL-4R inhibitor.
]]></description>
<dc:creator><![CDATA[ Shi, Y., Nolden, K., Ho, M., Li, H., Batista, V. S., Collazo, R., Bunick, C. G. ]]></dc:creator>
<dc:date>2026-04-13</dc:date>
<dc:identifier>doi:10.64898/2026.04.12.718052</dc:identifier>
<dc:title><![CDATA[Crystal structure and molecular dynamics simulations of rademikibart Fab-IL-4Rα complex reveal biochemical basis for next-generation potent IL-4Rα inhibition in type 2 allergic and inflammatory diseases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.09.717569v1?rss=1">
<title>
<![CDATA[
A novel sample delivery method for powder X-ray diffraction at Turkish Light Source 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.09.717569v1?rss=1
</link>
<description><![CDATA[
Powder X-ray diffraction is highly sensitive to sample-delivery conditions, particularly when measurements are performed on platforms originally designed for single-crystal diffraction. In this study, we developed a modified Terasaki-plate-based sample-delivery method for PXRD using a laboratory single-crystal diffractometer implemented with the XtalCheck-S plate-reader module at Turkish Light Source. The method was evaluated against standard loop/pin-based loading and a grease-based Terasaki setup using [4-(2-methoxyphenyl)piperazin-1-yl]methyl}-1,3,4-oxadiazol-2-thiol as a model analyte. While the loop-based method allowed initial PXRD measurements, it provided limited sample volume and insufficient particle statistics. The grease-based plate setup enabled multi-well data collection at a time, but yielded weaker, more diffuse patterns due to increased background noise. Inversely, modification of the Terasaki wells with Kapton tape enabled secure low-volume powder loading, improved diffraction clarity, and supported batch-mode data collection. Comparative search-match and profile-fitting analyses showed that all three loading strategies sampled the same crystalline material, whereas the Kapton-based setup presented the most reliable diffraction profile, with the lowest profile residual (Rp = 9.6%). These findings indicate that this novel sample-delivery method, rather than instrument hardware, can largely improve PXRD performance on an existing in-situ crystallography platform. The Kapton-Terasaki method provides a simple, cost-effective, and practical pipeline for high-throughput PXRD analysis of small powder samples under laboratory conditions.
]]></description>
<dc:creator><![CDATA[ AYAN, E., Kepceoglu, A., Mermer, A. ]]></dc:creator>
<dc:date>2026-04-13</dc:date>
<dc:identifier>doi:10.64898/2026.04.09.717569</dc:identifier>
<dc:title><![CDATA[A novel sample delivery method for powder X-ray diffraction at Turkish Light Source]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.12.718003v1?rss=1">
<title>
<![CDATA[
SLFN-generated 3'-truncated tRNALeu or tRNASer together with tRNase ZL works as a sequence-specific RNA cutter 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.12.718003v1?rss=1
</link>
<description><![CDATA[
A complex of the 3'-truncated tRNAArg that lacks 9 nt and tRNase ZL works as a GCCC-recognizing RNA cutter. It recognizes an RNA substrate via four Watson-Crick-Franklin base-pairings with the 3'-truncated tRNAArg. Human SLFN11 and SLFN13 can generate 3'-truncated tRNALeu that lacks 10 nt and 3'-truncated tRNASer that lacks 11 nt, respectively, from their corresponding mature tRNAs. Here, we investigated if these 3'-truncated tRNAs together with tRNase ZL work as sequence-specific RNA cutters. We examined five RNA targets for cleavage by recombinant human tRNase ZL in the presence of the 3'-truncated tRNALeu or tRNASer. We demonstrated that the 3'-truncated tRNALeu and tRNASer together with tRNase ZL indeed work as [~]6-base-recognizing and 7-base-recognizing RNA cutters, respectively.
]]></description>
<dc:creator><![CDATA[ Takahashi, M., Nashimoto, M. ]]></dc:creator>
<dc:date>2026-04-13</dc:date>
<dc:identifier>doi:10.64898/2026.04.12.718003</dc:identifier>
<dc:title><![CDATA[SLFN-generated 3'-truncated tRNALeu or tRNASer together with tRNase ZL works as a sequence-specific RNA cutter]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.09.717471v1?rss=1">
<title>
<![CDATA[
A Multi-Objective Scoring (MOS) Framework for Detecting Cross-Modal Spatial Similarity: Conceptual and Direct Formulations 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.09.717471v1?rss=1
</link>
<description><![CDATA[
Spatial multi-omics methods allow researchers to study complex biological systems by integrating multiple molecular layers while preserving their spatial organization. However, integrating spatial transcriptomics and mass spectrometry imaging (MSI) remains challenging due to the differences between the two modalities, including sampling geometry, spatial resolution, signal scaling, and measurement principles. For example, 10x Genomics Visium captures transcriptomic data on a discrete hexagonal grid of spots; however, matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) produces dense Cartesian pixel maps of molecular distributions. These differences make exact spatial co-registration difficult and limit the effectiveness of single-metric similarity approaches.

Here, we introduce a Multi-Objective Scoring (MOS) framework designed to detect cross-modal spatial similarity without requiring exact pixel-to-pixel alignment. The MOS framework integrates multiple complementary spatial descriptors into a unified similarity score, including coordinate-based metrics (value correlation, importance-based Intersection-over-Union (IoU), and importance-map correlation) and descriptor-based metrics that capture higher-order spatial organization, such as spatial histograms, radial profiles, quadrant statistics, and Morans I spatial autocorrelation. These metrics are combined through a weighted ensemble model. This model calculates the weights using synthetic spatial datasets that simulate realistic tissue geometry, sampling differences, and spatial distortions.

The framework was applied to a spatial multi-omics dataset from murine brain tissue, integrating spatial transcriptomics with MALDI-MSI lipidomics across young and aged control and Alzheimers disease (AD) models. Synthetic data validation results demonstrated strong pattern-matching performance (96.14% accuracy), and application to experimental data identified several MSI analyte features whose spatial distributions closely matched transcriptomic patterns. In particular, strong and reproducible associations were observed between myelin-related genes (Mbp and Plp1) and multiple analyte features enriched in white matter regions.

Overall, whether applied conceptually or directly, the MOS framework provides a strong strategy for cross-modal spatial integration and offers a scalable tool for discovering spatial relationships across diverse multi-omics datasets and facilitating hypothesis generation.
]]></description>
<dc:creator><![CDATA[ Jarrahi, A., Jones, A. R., Tang, W., Qi, H., Crouch, A. C. ]]></dc:creator>
<dc:date>2026-04-13</dc:date>
<dc:identifier>doi:10.64898/2026.04.09.717471</dc:identifier>
<dc:title><![CDATA[A Multi-Objective Scoring (MOS) Framework for Detecting Cross-Modal Spatial Similarity: Conceptual and Direct Formulations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.12.717905v1?rss=1">
<title>
<![CDATA[
Phosphorylation of UBE2J1 at serine residue S184 contributes towards infection and cellular syncytialization by Vesicular Stomatitis Virus 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.12.717905v1?rss=1
</link>
<description><![CDATA[
The ubiquitin-conjugating enzyme UBE2J1 functions in the proteasomal degradation of proteins at the ER. Existing evidence suggests that it plays a role during viral infection, with elevated UBE2J1 levels generally associated with increased infection. This is particularly relevant for some RNA viruses; however, the regulation of UBE2J1 during infection has not been well studied. Here, we used a BHK21 cell model to demonstrate that UBE2J1 overexpression promotes the replication of Vesicular Stomatitis Virus (VSV), as indicated by a significant increase in viral titres. To better understand the underlying molecular processes, cells were co-transfected to express the VSV-G protein and wild-type UBE2J1 protein, and we observed a significant increase in the syncytial fusion area. This effect was not observed when catalytically inactive (C91S) or phospho-deficient (S184A) forms of the protein were used. Interestingly, overexpression of a truncated, non-ER localized form of UBE2J1 ({Delta}TM) led to the largest increase in the syncytial fusion area. This arose as a result of increased syncytia size, and may indicate an enhanced cellular role if soluble forms of UBE2J1 are not anchored to the ER. Additional studies using truncated, mutated and wild-type proteins confirmed that UBE2J1 increases VSV viral replication and is associated with an increase in the number of infection plaques. Considering the emerging evidence for UBE2J1 involvement in viral infection, our finding should help in understanding the role of this protein in viral pathogenesis and cellular processes linked to syncytialization.
]]></description>
<dc:creator><![CDATA[ Algoufi, N. D., Walsh, E. B., Fallata, Z. I., Alamri, S. S., Hashem, A. M., Fleming, J. V. ]]></dc:creator>
<dc:date>2026-04-12</dc:date>
<dc:identifier>doi:10.64898/2026.04.12.717905</dc:identifier>
<dc:title><![CDATA[Phosphorylation of UBE2J1 at serine residue S184 contributes towards infection and cellular syncytialization by Vesicular Stomatitis Virus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.10.715585v1?rss=1">
<title>
<![CDATA[
Mechanism of GPR84 allosteric modulation at a helix 8-proximate site 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.10.715585v1?rss=1
</link>
<description><![CDATA[
Allosteric modulators offer opportunities for pathway-selective GPCR signalling, but the structural mechanisms enabling biased allosteric modulation remain unclear. Here we identify a helix 8-proximate allosteric site in the immune-metabolic receptor GPR84 and define how it achieves Gi -biased signalling. Cryo-EM structures of the GPR84-Gi complexes bound to the orthosteric agonist OX04539 alone or in combination with the positive allosteric modulator (PAM) PSB-16671 reveal that PSB-16671 binds at the interface of TM1, TM7, and helix 8, a location distinct from previously characterized GPCR allosteric pockets. Molecular dynamics simulations and mutagenesis uncover a polar interaction network linking orthosteric and allosteric sites through conserved residues including Asp662.50, Asn1043.36, and Asn3627.45. Unexpectedly, disrupting this network enhances allosteric cooperativity, indicating that conformational flexibility within the network is essential for allosteric communication. PSB-16671 stabilizes a receptor conformation with pronounced TM6 displacement that favours Gi coupling while disfavouring {beta}-arrestin recruitment. This Gi-biased profile sustains macrophage phagocytosis of cancer cells without the desensitization induced by balanced agonists. Sequence analysis suggests that helix 8-proximate allosteric sites may be broadly targetable across class A GPCRs, while receptor-specific contacts enable selective modulation. These findings establish structural and mechanistic principles for biased allosteric modulation applicable beyond GPR84.
]]></description>
<dc:creator><![CDATA[ Zhang, X., Guseinov, A.-A., Jenkins, L., Zhou, J., Gossen, F., Wang, P., Al Mahmud, Z., Li, Y., Mahardhika, A. B., Muller, C. E., Feng, M., Russell, A. J., Tikhonova, I. G., Milligan, G., Zhang, C. ]]></dc:creator>
<dc:date>2026-04-12</dc:date>
<dc:identifier>doi:10.64898/2026.04.10.715585</dc:identifier>
<dc:title><![CDATA[Mechanism of GPR84 allosteric modulation at a helix 8-proximate site]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.09.717382v1?rss=1">
<title>
<![CDATA[
Beyond the surface: plasmalogens are dispensable for retinal integrity and fertility in the mouse 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.09.717382v1?rss=1
</link>
<description><![CDATA[
Ether lipids and their subclass, the plasmalogens, are critical regulators of membrane organization, signaling, and stress responses in multiple tissues. Inborn errors in their anabolism cause severe multi-organ diseases such as Rhizomelic Chondrodysplasia Punctata and related peroxisomal disorders. The Gnpat knockout mouse model, characterized by total ether lipid deficiency, recapitulates key features of this disorder, including dense bilateral cataracts, microphthalmia, and infertility, but the specific contribution of different subclasses like plasmalogens remains elusive. The recent identification of the Peds1 gene allows dissecting the impact of selective plasmalogen deficiency with retention of plasmanyl lipids, another ether lipid subgroup. Here, we performed the first side-by-side comparison of Gnpat and Peds1 knockout in mice on a matched genetic background (C57BL/6 x CD1). In contrast to the situation in Gnpat knockout mice, plasmanyl lipids in Peds1 knockout mice were sufficient to prevent cataract formation and maintain normal ocular structures, despite marked shifts in the ocular phospholipidome. Also, fertility and reproductive function were found to be preserved in Peds1 knockout mice. Our data demonstrate that plasmanyl lipids can partially protect against the severe phenotypes observed in mouse models of total ether lipid deficiency; notably, the ocular and reproductive phenotypes were plasmalogen-independent, indicating that loss of the vinyl ether double bond is not the key determinant of all symptoms in human and murine ether lipid deficiency and can at least partly be compensated by plasmanyl lipids.

HighlightsO_LIIn mice, total ether lipid deficiency causes cataracts and infertility.
C_LIO_LIThe role of plasmalogens in these phenomena remains unclear.
C_LIO_LITwo PEDS1-deficient patients were reported, but cataracts were observed in only one case.
C_LIO_LIPeds1-deficient mice have no cataracts or ocular abnormalities.
C_LIO_LIMice with a deletion of Peds1 display normal fertility rates.
C_LI
]]></description>
<dc:creator><![CDATA[ Dorigatti, I., Juric, V., Blumer, M. J., Kummer, D., Kokot, J., Golderer, G., Dorninger, F., Berger, J., Keller, M. A., Watschinger, K. ]]></dc:creator>
<dc:date>2026-04-12</dc:date>
<dc:identifier>doi:10.64898/2026.04.09.717382</dc:identifier>
<dc:title><![CDATA[Beyond the surface: plasmalogens are dispensable for retinal integrity and fertility in the mouse]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.11.717886v1?rss=1">
<title>
<![CDATA[
Molecular dissection of protein complexes isolated from sections of human brain 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.11.717886v1?rss=1
</link>
<description><![CDATA[
Molecular studies of brain receptors and transporters have typically relied on recombinant systems, limiting insight into their organization in native tissue. Here, we develop nanobody-based immunoprecipitation coupled with native mass spectrometry to interrogate endogenous protein assemblies from post-mortem mouse and human brain sections. We exemplify our approach by characterizing the synaptic proteins VGluT1 and mGluR2. From a single mouse brain, we discover mGluR2/3 heterodimers, alongside mGluR2 homodimers. Considering regions of human brain heterodimeric mGluR2/3 is highly abundant in the OFC and sgACC ([~]70% and 50%, respectively) and forms regional-specific interactions with additional synaptic proteins. In a modest cohort of biobanked human tissue, associated with depression and suicide, we find increased mGluR2/3 in the OFC. Consistent with this, mice exhibit similar associations between heterodimer levels and stress-susceptibility. Overall, our approach provides a direct means for establishing molecular-behavioural links at the level of receptor organization in brain.
]]></description>
<dc:creator><![CDATA[ El-Baba, T. J., Lutomski, C. A., Bennett, J. L., Lawrence, S. A. S., Burnap, S. A., Butroid, F. I., Ramsay, O. B., Radzevicius, T., Wu, D., Song, H., Chan, K. L., Parise, L. F., Parise, E., Struwe, W. B., Murrough, J. W., Russo, S. B., Robinson, C. V. ]]></dc:creator>
<dc:date>2026-04-12</dc:date>
<dc:identifier>doi:10.64898/2026.04.11.717886</dc:identifier>
<dc:title><![CDATA[Molecular dissection of protein complexes isolated from sections of human brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.04.09.717454v1?rss=1">
<title>
<![CDATA[
Proteomics-Based Discovery of Symmetry-Specific Readers and Antireaders of 5-Formylcytosine in Mammalian DNA 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.04.09.717454v1?rss=1
</link>
<description><![CDATA[
5-methylcytosine (mC) and its oxidized derivatives are epigenetic modifications of mammalian DNA that play key roles in transcription, cell differentiation, and cancer. They predominantly occur within palindromic CpG dyads, creating multiple possible combinations across the two dyad strands, each representing a chemically distinct DNA major groove mark. Among the modifications, 5-formylcytosine (fC) interacts with a large number of proteins and has been associated with important roles in chromatin regulation. However, it is poorly understood whether nuclear proteins selectively recognize the symmetry of individual fC-containing CpG dyads. Here, we report the first proteome-wide interaction profiles of symmetric and asymmetric fC modifications in mammalian DNA. Our analysis spans several human and mouse cell lines and three DNA promoter probes containing CpG dyads with five distinct combinations of fC, C, and mC nucleobases. We identify a diverse set of fC reader proteins with distinct symmetry preferences, including transcription factors (e.g., MAX, HEY1, RFX5, SIX1, SIX2, and FOXJ3) and DNA repair proteins (MPG and TDG). Notably, some proteins act as fC readers or anti-readers depending on the sequence context, suggesting a role for fC in modulating their target selection. Our data reveal widespread symmetry-specific recognition of fC by mammalian proteins and provide a rich resource for studying its potential functions in chromatin regulation.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=183 HEIGHT=200 SRC="FIGDIR/small/717454v1_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@1735842org.highwire.dtl.DTLVardef@1a7914borg.highwire.dtl.DTLVardef@340817org.highwire.dtl.DTLVardef@1ff33a6_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator><![CDATA[ Cakil, Z. V., Engelhard, L., Seidel, N., Eppmann, S., Bange, T., Summerer, D. ]]></dc:creator>
<dc:date>2026-04-12</dc:date>
<dc:identifier>doi:10.64898/2026.04.09.717454</dc:identifier>
<dc:title><![CDATA[Proteomics-Based Discovery of Symmetry-Specific Readers and Antireaders of 5-Formylcytosine in Mammalian DNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
