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<title>bioRxiv Subject Collection: Biophysics</title>
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<description>
This feed contains articles for bioRxiv Subject Collection "Biophysics"
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<title>bioRxiv</title>
<url>https://www.biorxiv.org/sites/default/files/bioRxiv_article.jpg</url>
<link>https://www.biorxiv.org</link>
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<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.14.724942v1?rss=1">
<title>
<![CDATA[
Towards a database capturing chromosome structure and function: symbols and syntax 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.14.724942v1?rss=1
</link>
<description><![CDATA[
Existing databases of interphase chromosome conformations typically store three-dimensional coordinates of genomic segments. However, since interphase chromatin is highly dynamic, such databases are dominated by transient configurations and unstructured regions, whose positions vary continuously between cells and over time, unlike folded proteins such as globin, which adopt similar structures in every cell. These drawbacks motivated the inception of a database based on 'strion' (a portmanteau of a string capturing structure and function). A strion concisely describes the structure and activity of all transcription units in one cell, by retaining only functionally relevant positional information. Sets of strions describing structures in different cells sampled at different times are compiled into a 'super-strion'. Then, 46 super-strions summarise the range of structure and activity of a human cell type, including information on all transcription units, how often each co-fires and co-clusters with others in transcription factories/hubs, enhancer interactomes and small-world expression networks.
]]></description>
<dc:creator><![CDATA[ Cook, P. R., Marenduzzo, D., Valei, Z. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.14.724942</dc:identifier>
<dc:title><![CDATA[Towards a database capturing chromosome structure and function: symbols and syntax]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.13.724817v1?rss=1">
<title>
<![CDATA[
pH Induced Changes in Protein Structure and Hydration 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.13.724817v1?rss=1
</link>
<description><![CDATA[
The molten globule (MG) state is an intermediate in the unfolding pathway of proteins, typically triggered by denaturing agents such as urea, extreme pH, high pressure, or heat. The microscopic details of such states are far from understood. Here we study the MG states in protein Hen Egg-White Lysozyme (PDB ID: 1AKI) using microscopic constant pH molecular dynamics (CpHMD) simulations and experiments across a wide pH range. We observe that the titratable residues act as key drivers of conforma- tional fluctuations, promoting the emergence of MG states at extreme pH. These states display partial unfolding, and small global structural changes (< 7% deviation). Hydration around the fluctuating acidic residues shows reduced water density and weakened hydrogen bonding at low pH. At high pH, hydration around acidic residues increases relative to pH = 7, whereas hydration around basic residues decreases. The translational and rotational dynamics of hydration water also exhibit pronounced pH dependence: the translational diffusion coefficient (Dtrans) increases linearly with decrease in pH in acidic medium and increases linearly with increasing pH in the basic regime. The rotational diffusion (Drot) shows similar dependencies on pH except a break at pH {approx} 4 corresponding to acidic residue pKa values. Our results may be useful to identify ligand binding of lysozyme in extreme pH conditions.
]]></description>
<dc:creator><![CDATA[ Sen, A., Chakrabarti, J., Mitra, R. K. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.13.724817</dc:identifier>
<dc:title><![CDATA[pH Induced Changes in Protein Structure and Hydration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.11.724350v1?rss=1">
<title>
<![CDATA[
Cryo-EM structure and biochemical characterization of a BRAF/CRAF heterodimer: Negative charge in the NtA motif is not required for RAF activation 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.11.724350v1?rss=1
</link>
<description><![CDATA[
Abstract Upon RAS-driven membrane recruitment, RAF kinases ARAF, BRAF and CRAF are activated via formation of homo- or hetero- dimers to initiate signaling through the MAP kinase cascade. Although RAF heterodimers are important for both physiologic and oncogenic signaling, they have been little studied at a structural and biochemical level. Here we report the preparation, biochemical characterization, and the cryo-EM structure of a 14-3-3-bound BRAF/CRAF heterodimer complex. The heterodimer exhibited kinetic parameters and sensitivity to a panel of twelve structurally diverse RAF inhibitors that were closely similar to, or intermediate between, those of BRAF and CRAF homodimers. Cryo-EM structures of the heterodimer with and without MEK1 revealed an overall organization essentially identical to that of RAF homodimers, but with an asymmetric interaction in the MEK1-bound structure in which the BRAF N-terminal acidic (NtA) motif extends across the dimer interface to engage the CRAF RKTR motif. Mutagenesis of this interface unexpectedly revealed that replacing the acidic NtA sequence with a basic RARA sequence yields highly active RAF homodimers and heterodimers, demonstrating that negative charge in the NtA motif is not required for activity. Collectively, our findings suggest that the charge state of the NtA motif influences RAF activity through effects on local backbone dynamics and the stability of the inactive kinase conformation, rather than via stereospecific recognition across the dimer interface.
]]></description>
<dc:creator><![CDATA[ Ha, B. H., Tkacik, E., Gazgalis, D., Kang, H., Jang, D. M., Chakraborty, S., Jeon, H., Eck, M. J. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.11.724350</dc:identifier>
<dc:title><![CDATA[Cryo-EM structure and biochemical characterization of a BRAF/CRAF heterodimer: Negative charge in the NtA motif is not required for RAF activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.11.724279v1?rss=1">
<title>
<![CDATA[
Structural basis for the intestinal protocadherin-based intermicrovillar adhesion complex 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.11.724279v1?rss=1
</link>
<description><![CDATA[
The intestinal brush border (BB), composed of densely packed microvilli on enterocytes, is essential for nutrient absorption and host defense. Its organization relies on the intermicrovillar adhesion complex (IMAC), mediated by protocadherins CDHR2 and CDHR5. Despite their clinical relevance in inflammatory bowel disease and several carcinomas, structural details of IMAC assemblies have remained elusive. Herein, we report the Cryo-EM structure of the adhesive complex at 3.4 [A] resolution, revealing a heterotetrameric ensemble composed of a dimer of CDHR2 and a dimer of CDHR5. This assembly ensures uniform adhesive strength between neighboring microvilli, and facilitates hexagonal packing of microvilli. Biophysical analyses and molecular dynamics simulations revealed a kinked, Ca2+-free linker between domains EC3 and EC4 of CDHR5 conferring the necessary flexibility to withstand the shear stress caused during intestinal peristalsis. Collectively, these findings provide a structural framework for understanding BB organization and suggest strategies for therapeutics targeting IMAC in intestinal disorders.
]]></description>
<dc:creator><![CDATA[ Senoo, A., Guillen-Poza, P., Fujishima, K., Kosuge, H., Doumoto, T., Kasahara, K., Tanihara, T., Yoshida, Y., Yanaka, S., Nakakido, M., Nagatoishi, S., Maenaka, K., Ohdo, S., Matsunaga, N., Hervas, R., Tsumoto, K., Caaveiro, J. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.11.724279</dc:identifier>
<dc:title><![CDATA[Structural basis for the intestinal protocadherin-based intermicrovillar adhesion complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.11.724287v1?rss=1">
<title>
<![CDATA[
RNA synthesis and degradation regulate biomolecular condensates through non-equilibrium feedback 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.11.724287v1?rss=1
</link>
<description><![CDATA[
Transcriptional condensates operate far from equilibrium, where continuous RNA synthesis and degradation dynamically reshape condensate composition. To investigate how RNA synthesis regulates condensate properties at sub-molecular resolution, we introduce REACT-RNA, a chemically specific coarse-grained molecular dynamics framework that explicitly couples RNA polymerisation, degradation, and nucleotide fluxes to sequence-dependent protein-RNA phase behaviour. Using FUS and MED1 as model systems, we show that RNA growth remodels condensate phase behaviour by altering RNA length distributions and intermolecular connectivity. Sustained RNA polymerisation drives re-entrant condensate dissolution, even of aged gel-like condensates, whereas RNA degradation stabilises long-lived non-equilibrium condensates containing excess RNA and negative charge beyond that tolerated at equilibrium. Our results suggest that RNA synthesis, degradation, and nucleotide fluxes drive transcriptional condensates out of thermodynamic equilibrium while condensates in turn promote reactive molecular configurations that favour RNA production, enabling transient accumulation of excess RNA and negative charge beyond equilibrium electroneutrality constraints during bursts of transcription.
]]></description>
<dc:creator><![CDATA[ Sanchez-Burgos, I., Tejedor, A. R., Ocana, A., R. Espinosa, J., Collepardo-Guevara, R. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.11.724287</dc:identifier>
<dc:title><![CDATA[RNA synthesis and degradation regulate biomolecular condensates through non-equilibrium feedback]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.11.724295v1?rss=1">
<title>
<![CDATA[
Large-domain histology-based diffusion MRI simulation via independent local simulations 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.11.724295v1?rss=1
</link>
<description><![CDATA[
Diffusion MRI simulations based on realistic tissue microstructure provide a means to validate biophysical models and optimize acquisition protocols, but their computational cost restricts most studies to domains far smaller than a clinical voxel. The objective of this study was to develop an automated and scalable framework that converts whole-slide histology into diffusion MRI simulations at clinically relevant spatial scales while remaining feasible on standard workstation hardware. We present an end-to-end pipeline integrating two-dimensional whole-slide cell segmentation, mesh generation, and finite element Bloch-Torrey simulation. To enable simulations at large spatial scales without prohibitive memory growth, we introduce a subdomain tiling strategy in which the tissue domain is partitioned into extended subdomains simulated independently under no-flux boundary conditions. Signals are aggregated only from the central regions of each subdomain to minimize boundary artifacts. For an 800 m x 800 m histology-based domain, the aggregated signal differed by 0.07% from the corresponding full-domain finite element simulation while reducing wall-clock time from several days to hours and maintaining bounded memory usage independent of global domain size. When applied to a 2016 m x 2016 m heterogeneous region approximating the in-plane dimensions of a clinical voxel, the apparent diffusion coefficient obtained from the full domain differed from values computed in smaller dense and sparse subregions, demonstrating the influence of structural heterogeneity at clinically relevant scales on derived diffusion metrics. The proposed framework establishes an automated and memory-stable approach for generating diffusion MRI simulations directly from routine histology.
]]></description>
<dc:creator><![CDATA[ Kohler, I. A., Zheng, L., Kuder, T. A., Goedicke, O., Ladd, M. E., Hesser, J. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.11.724295</dc:identifier>
<dc:title><![CDATA[Large-domain histology-based diffusion MRI simulation via independent local simulations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.11.724289v1?rss=1">
<title>
<![CDATA[
PolyA/polyQ-mediated conformational rewiring regulates DNA engagement and drives aggregation in the neuronal transcription factor Ascl1 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.11.724289v1?rss=1
</link>
<description><![CDATA[
Ascl1 is a pioneer transcription factor that drives neuronal fate decisions, yet the structural basis of its activity remains elusive. Besides the basic helix-loop-helix (bHLH) domain which dimerizes with other transcription factors and binds DNA, Ascl1 contains long low-complexity intrinsically disordered regions (IDRs), including a polyA/polyQ tract of unknown function. Here we apply single-molecule FRET to generate a conformational map of full-length Ascl1 across different functional states that reveals how the polyA/polyQ tract finely tunes molecular properties. Monomeric Ascl1 is largely disordered but displays sensitive interdomain coupling between the N-terminal IDR and the bHLH domain where the polyA/polyQ tract destabilizes the latter. Heterodimerization with the transcription factor E12 not only promotes folding of the bHLH domain and high-affinity DNA binding but also relieves interdomain interactions including the low complexity polyA/polyQ tract, increasing both its extension and chain dynamics. Remarkably, deletion of the polyA/polyQ tract not only dramatically reduces non-specific DNA binding but also abolishes aggregation in vitro and increases Ascl1 abundance in human HEK293T cells. Overall, our results highlight the capacity of intrinsically disordered and low-complexity regions to impart diverse regulatory roles in transcription factors.
]]></description>
<dc:creator><![CDATA[ Mitra, S., Ruidiaz, S. F., Laufeyjardottir, A., Voutsinos, V., Nitsenko, K., Guala, A., Zaret, K. S., Hartmann-Petersen, R., Heidarsson, P. O. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.11.724289</dc:identifier>
<dc:title><![CDATA[PolyA/polyQ-mediated conformational rewiring regulates DNA engagement and drives aggregation in the neuronal transcription factor Ascl1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.12.724662v1?rss=1">
<title>
<![CDATA[
Membrane Phase, Charge, and Curvature Regulate α-Synuclein Binding Dynamics 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.12.724662v1?rss=1
</link>
<description><![CDATA[
-Synuclein (Syn) is an intrinsically disordered protein whose interactions with lipid membranes are central to both its physiological function and its role in synucleopathies. While membrane charge, phase, and curvature are each known to influence Syn binding, these properties are typically examined independently, leaving their combined effects on both equilibrium and dynamic membrane association unresolved. Here, we systematically investigate how membrane phase and charge jointly regulate Syn binding, curvature sensitivity, and exchange dynamics using fluorescence microscopy, circular dichroism spectroscopy, and fluorescence recovery after photobleaching (FRAP), complemented by coarse-grained molecular dynamics simulations. Under zwitterionic conditions, Syn preferentially binds highly curved gel-phase membranes, driven by curvature-dependent enrichment of packing defects arising from faceted vesicle morphologies. Incorporation of anionic lipids selectively enhances binding in liquid-phase membranes while attenuating curvature-dependent partitioning in gel-phase membranes. Dynamic measurements reveal that membrane phase and charge also govern the stability of membrane-associated Syn, with gel-phase membranes and anionic lipids promoting kinetically stabilized states. Simulations show that curvature-induced defect formation is strongly amplified in gel-phase membranes but largely insensitive to charge. These findings establish that Syn-membrane interactions are governed by a cooperative interplay between membrane phase, curvature, and charge and highlight the importance of resolving both thermodynamic and kinetic contributions to protein-membrane binding.
]]></description>
<dc:creator><![CDATA[ Kou, O. H., Sakurai, C. M., Ramirez, S. Y., Kim, B. H., Johnson, D. H., Zhang, Z., Lee, C. T., Zeno, W. F. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.12.724662</dc:identifier>
<dc:title><![CDATA[Membrane Phase, Charge, and Curvature Regulate α-Synuclein Binding Dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.12.724610v1?rss=1">
<title>
<![CDATA[
Substrate mediated mechanical forces enable optimal kinetic proofreading by T-cell receptors 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.12.724610v1?rss=1
</link>
<description><![CDATA[
T-cells use molecular reactions with nonequilibrium error correction, i.e., proofreading, to discriminate between nearly identical antigens with high specificity and sensitivity. These receptor binding events are known to be force sensitive, yet traditional schemes of proofreading focus on reaction kinetics alone and do not consider the role of force dependent catch/slip bond behavior or interactions with mechanically engaged coreceptors such as adhesion molecules. To address this, we propose a minimal framework for proofreading of ligand discrimination by T-cell receptors (TCRs) that uses endogenous TCR mechanosensation and substrate-mediated mechanical interactions with adhesive proteins (load sharing) to improve recognition fidelity. We leverage the catch bond behavior of cognate antigens to delay decision making and amplify TCR signaling while discarding noncognate slip bond ligands in the presence of a force. By integrating our model with existing structural and molecular data, we show that substrate mechanics regulates the transmission of active cytoskeletal forces through a molecular clutch and controls the energization of bound TCRs needed for optimal proofreading. Our work demonstrates how mechanical forces and substrate properties can augment kinetic proofreading in T-cells, suggesting biomaterial design strategies for immunotherapies that tune the mechanical microenvironment of T-cells to achieve high fidelity TCR-ligand discrimination, antigen recognition, and activation.
]]></description>
<dc:creator><![CDATA[ Jeffreys, N., Brockman, J. M., Heydari, T., Nerger, B. A., Jung, W.-H., Zandstra, P. w., Mahadevan, L., Mooney, D. J., Shankar, S. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.12.724610</dc:identifier>
<dc:title><![CDATA[Substrate mediated mechanical forces enable optimal kinetic proofreading by T-cell receptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.13.724476v1?rss=1">
<title>
<![CDATA[
Fungal Hyphae as Distributed Vapor Sinks 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.13.724476v1?rss=1
</link>
<description><![CDATA[
Hygroscopic surfaces act as local vapor sinks that reshape the condensation field around them, but whether distributed biological structures do the same has not been investigated. We have established that hyphae of fungal colonies functionally be have as vapor sinks, creating a dry region of width {delta} around themselves when placed on a cooled substrate. In addition, the radial distribution of droplet sizes steepens during condensation, and the rate at which droplets evaporate locally after chamber drying increases. In order to quantify this behavior, we employed a combination of time-resolved imaging and survival analysis to determine how long individual droplets persist on the surface surrounding the colony. These data were used to derive three quantitative measures of the vapor-sink effect. Each measure was found to be directly proportional to the vapor-sink strength of the substrate, as calibrated against NaCl-agar hydrogels of known water activity (LOOCV RMSE = 0.031 for recovered aw). These findings were consistent across three fungal genera (35 experiments), and all species fell along calibration lines defined by the hydrogel standards. This result is consistent with a diffusion-limited vapor-depletion framework. The measured genus-level {delta} ratios agreed to within 6% of predictions from structural absorbing capacity, and field measurements on Gymnosporangium-infected apple leaves were consistent with the same signatures under natural conditions. These results establish a non-contact method for inferring the material properties of thin hygroscopic biological surfaces from their condensation patterns.
]]></description>
<dc:creator><![CDATA[ Lin, Y. J., Feng, L., Khan, A., Park, K.-c., Jung, S. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.13.724476</dc:identifier>
<dc:title><![CDATA[Fungal Hyphae as Distributed Vapor Sinks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.12.724524v1?rss=1">
<title>
<![CDATA[
A workflow for the identification of oligomeric structures on tilted sample planes in Cryo-SMLM 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.12.724524v1?rss=1
</link>
<description><![CDATA[
We introduce a workflow to identify oligomeric structures that are recorded with single-molecule localization microscopy (SMLM) under cryogenic conditions. Typically, these oligomers are assumed to consist of protomers arranged as equilateral two-dimensional polygons and every protomer is labeled with a dye molecule for visualization. Unlike previous work, we consider scenarios in which the sample plane has an unknown orientation relative to the focal plane. Our contribution is a high-precision plane-fitting algorithm to determine the sample plane, combined with geometrical transformations and two circle-fitting algorithms to identify the oligomeric structures. Our simulations on synthetic data demonstrate that the proposed workflow achieves high accuracy in estimating both the unknown tilted plane and the oligomer size.
]]></description>
<dc:creator><![CDATA[ Dong, Y., Yang, Z., Schneider, M., Scherzer, O., Schuetz, G. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.12.724524</dc:identifier>
<dc:title><![CDATA[A workflow for the identification of oligomeric structures on tilted sample planes in Cryo-SMLM]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.13.724805v1?rss=1">
<title>
<![CDATA[
Temperature-dependent ligand relocation reveals plasticity of TRPM4 inhibition 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.13.724805v1?rss=1
</link>
<description><![CDATA[
Transient receptor potential melastatin 4 (TRPM4) is a Ca2+-activated cation channel whose pharmacology is shaped by its molecular environment. It remains poorly understood how temperature and membrane context influence inhibitor recognition. Here we combine cryo-electron microscopy of membrane-derived vesicles and detergent-solubilized TRPM4 to investigate lipid-associated architecture and binding of the potent anthranilic anilide inhibitor PBA. We find that membrane vesicles preserve a native-like paralipid environment and reveal lipid binding patterns highly similar to those observed in GDN, supporting detergent-solubilized TRPM4 as a structurally relevant system for ligand analysis. Strikingly, PBA occupies distinct binding pockets at 8 {degrees}C and 37 {degrees}C. At low temperature, PBA binds in a previously described inhibitor pocket formed by S3, S4, the S4-S5 linker and the TRP helix, whereas at physiological temperature it relocates to a distinct site within the S1-S4 domain proximal to the Ca2+ regulatory region. These findings reveal temperature-dependent plasticity in TRPM4 ligand recognition.
]]></description>
<dc:creator><![CDATA[ Schneiter, D. M., Rougier, J.-S., Abriel, H., Stahlberg, H., Ekundayo, B. E. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.13.724805</dc:identifier>
<dc:title><![CDATA[Temperature-dependent ligand relocation reveals plasticity of TRPM4 inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.12.724695v1?rss=1">
<title>
<![CDATA[
Transthyretin amyloid fibrils adopt distinct folds in the brain 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.12.724695v1?rss=1
</link>
<description><![CDATA[
Amyloid deposition in the central nervous system is increasingly recognized in transthyretin (ATTR) amyloidosis, particularly in patients with prolonged survival following liver transplantation or disease-modifying therapies. However, the structural basis of transthyretin aggregation in the brain remains unknown. Here we determine cryo-electron microscopy (cryo-EM) structures of ex vivo brain-derived ATTR fibrils from patients carrying the ATTRv-V30M and ATTRv-V30G variants. Both fibrils adopt folds distinct from those previously reported in peripheral tissues and the vitreous humor. V30M fibrils exhibit a continuous ordered core spanning residues Pro11-Asn124, whereas V30G fibrils consist of a substantially reduced ordered core, revealing pronounced structural divergence even within the same tissue environment. Despite this diversity, comparative analyses identify conserved regions across ATTR fibrils, including a segment implicated in transthyretin aggregation and targeted for diagnostic and therapeutic development. These results provide direct evidence that local tissue context can shape amyloid fibril architecture in human disease.
]]></description>
<dc:creator><![CDATA[ Saelices, L., Afrin, S., Nguyen, B. A., Bassett, P. T., Fernandez Ramirez, M. d. C., Pedretti, R., Villalon, L., Kelly, C., Lopez, C., Madabushi, M., Zhou, A., Reis, I., Taipa, R., Evers, B. M. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.12.724695</dc:identifier>
<dc:title><![CDATA[Transthyretin amyloid fibrils adopt distinct folds in the brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.12.724125v1?rss=1">
<title>
<![CDATA[
Cellular Mechanisms of Transcranial Magnetic Stimulation in Climbing Fibers and Purkinje Neurons in the Cerebellum 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.12.724125v1?rss=1
</link>
<description><![CDATA[
Although transcranial magnetic stimulation (TMS) is widely used for brain stimulation, fundamental issues about its underlying mechanisms remain unresolved. We investigated some of these issues experimentally using an intact isolated turtle cerebellum in vitro, employing a novel chamber designed to deliver precisely calibrated induced electric fields along cortical depth. Our results show that single-pulse TMS can directly activate Purkinje cells and climbing fibers, and synaptically activate Purkinje cells via climbing fibers -- all within the first 1.2 ms. Specifically, current source density analysis showed that TMS directly (non-synaptically) activated (1) climbing fibers near the bend with intracellular current directed toward the axonal terminals and (2) Purkinje cells directly near the axon initial segment with intracellular current directed toward the distal dendrites. The thresholds for direct activation of climbing fibers and Purkinje cells were found to be very similar, 25 {+/-} 1 V/m. The climbing fibers synaptically activated Purkinje cells, as expected, with intracellular current originating in the proximal dendritic trunk and directed toward the distal dendrites. At higher electric fields (> 58 {+/-} 17 V/m), TMS synaptically activated dendritic currents in Purkinje cells. These results provide new insight into how TMS may activate afferent fibers and cell bodies of cortical neurons.
]]></description>
<dc:creator><![CDATA[ Okada, Y., Dong, C., Makaroff, S., Sundaram, P. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.12.724125</dc:identifier>
<dc:title><![CDATA[Cellular Mechanisms of Transcranial Magnetic Stimulation in Climbing Fibers and Purkinje Neurons in the Cerebellum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.10.724166v1?rss=1">
<title>
<![CDATA[
Kaiso reads methylated CpGs at nucleosome entry/exit and displaces the H3 tail 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.10.724166v1?rss=1
</link>
<description><![CDATA[
The zinc finger transcription factor Kaiso recognizes methylated CpG dinucleotides at silenced promoters and imprinted loci, but how it engages methylated DNA within the nucleosome remains unclear. To address this, we developed a DNMT1-based strategy for preparing site-specifically methylated nucleosomes with defined position and methylation state of the Kaiso recognition motif. Electrophoretic mobility shift assays show that Kaiso binds methylated nucleosomes with strong positional preference, with high-affinity engagement at the entry/exit site (SHL 6.5; Kd {approx} 100 nM), reduced affinity at SHL 5.5 (Kd {approx} 170 nM), and no methylation-dependent enhancement at dyad-proximal positions. Hemi- and fully methylated substrates bind Kaiso comparably at SHL 6.5, and the E535A mutation, which disrupts a key methyl-CpG contact, reduces binding in a methylation- and position-dependent manner. Solution NMR titrations of 15N-labeled H3 nucleosomes reveal that Kaiso binding perturbs a discrete set of H3 N-terminal tail residues, with chemical shifts trending toward free-peptide values, indicating release of the tail from its nucleosomal DNA contacts. This pattern closely resembles that produced by the pioneer factor Sox2 at the same nucleosomal region, suggesting H3 tail displacement is a general consequence of factor engagement at the nucleosome edge, independent of DNA-recognition mode. These results establish Kaiso as an active reader of methylated nucleosomal DNA that may prime local chromatin by exposing the H3 tail.
]]></description>
<dc:creator><![CDATA[ Malaga Gadea, F. C., Nikolova, E. N. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.10.724166</dc:identifier>
<dc:title><![CDATA[Kaiso reads methylated CpGs at nucleosome entry/exit and displaces the H3 tail]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.10.724101v1?rss=1">
<title>
<![CDATA[
Benchmarking generative AI and physics based molecular simulation for sampling conformational heterogeneity in T4 Lysozyme 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.10.724101v1?rss=1
</link>
<description><![CDATA[
Wild-type T4 lysozyme (T4L) is used as a benchmark to evaluate conformational sampling across generative AI, AI-accelerated molecular simulation (AMS), and physics-based enhanced molecular dynamics (EMD). A four-state model: exposed/open, exposed/closed, buried/open, and buried/closed; is defined using physically meaningful collective variables. While generative AI methods (AF-cluster, MSA subsampling of AlphaFold2, ConforFold, AlphaFlow, ESMFlow, ConfRover, BioEmu) largely sample only the exposed/open state, AMS integrating generative ensembles with iterative molecular dynamics, recovering all states and reproducing equilibrium populations similar to EMD and experimental smFRET signatures.
]]></description>
<dc:creator><![CDATA[ Bhakat, S. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.10.724101</dc:identifier>
<dc:title><![CDATA[Benchmarking generative AI and physics based molecular simulation for sampling conformational heterogeneity in T4 Lysozyme]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.10.723887v1?rss=1">
<title>
<![CDATA[
GluDs are ionotropic dopamine receptors tuned by G-proteins 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.10.723887v1?rss=1
</link>
<description><![CDATA[
Dopamine is a neurotransmitter essential for cognition, and its dysregulation is associated with neurological diseases. Historically, dopamine has been understood to signal exclusively through metabotropic receptors. Delta-type ionotropic glutamate receptors (GluDs), which have recently been established as ligand-gated ion channels, are fundamental for synaptic maintenance, are implicated in neurological disorders, and co-localize with dopaminergic machinery. Here, we report that dopamine is a direct agonist of GluDs, eliciting ionotropic activity, as visualized by cryo-electron microscopy (cryo-EM), bilayer recordings, mutagenesis, and patch clamp recordings. Dopamine binds to the GluD ligand binding domain, inducing clamshell closure and channel activation through a distinct molecular interface. GluD channel activity is tightly regulated by G-proteins, which act as molecular switches to tune GluD activity: free G{beta}{gamma} inhibits ligand-gating, while G or inactive G-protein heterotrimers enable dopamine-induced GluD currents. This tuning of GluD activity by G-proteins is uncoupled in a point mutation associated with neurodegeneration. These findings expand mechanisms of neuronal dopaminergic signaling, uncover how G-proteins tune GluD channel activity, and provide a framework for targeting GluDs in neurological diseases.
]]></description>
<dc:creator><![CDATA[ Wang, H., Weaver, M. G., Carrillo, E., Zheng, I., Gu, W., Khau, J., Mondal, A. K., Yanez, A., O'Brien, E. S., Jayaraman, V., Twomey, E. C. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.10.723887</dc:identifier>
<dc:title><![CDATA[GluDs are ionotropic dopamine receptors tuned by G-proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.10.724087v1?rss=1">
<title>
<![CDATA[
The Conformation of the Complementary Strand and the Deformation of the DNA Groove upon DDB2 Binding Justifies the Different Repair Rates for Cyclobutane Pyrimidine Dimers. 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.10.724087v1?rss=1
</link>
<description><![CDATA[
The repair of photo-induced DNA lesions through nucleotide excision repair machinery is still the source of important questions. It has been observed that the repair rate of the different cyclobutane pyrimidine dimers, i.e. the photoproducts induced by dimerization of two pi-stacked pyrimidines (T<>T, T<>C, C<>T, C<>C), depends on the nucleobases involved in the lesion. TT derivatives (T<>T) are removed more slowly than those containing cytosine, especially in 5'. Using all-atom molecular dynamics simulations and free-energy calculations, we demonstrate that the variation of the repair rate observed in human skin and in cultured cutaneous cell is associated to the recognition of the four lesions by the DDB2 protein moiety, and more specifically by the differential structural deformation induced on the complementary strand. Indeed, while C<>C and C<>T induce a larger deviation on the groove parameters, T<>T and T<>C, instead, affect DNA structure to a lesser extent. less affected. These effects then hamper differentially the downstream recruitment of the repair complexes. The observed DNA deformation correlates with the experimental repair rate and provides a structural rationale for the different repair rates of CPD by nucleotide excision repair machinery.
]]></description>
<dc:creator><![CDATA[ Kedjar, Y., Hognon, C., Douki, T., Dumont, E., MONARI, A. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.10.724087</dc:identifier>
<dc:title><![CDATA[The Conformation of the Complementary Strand and the Deformation of the DNA Groove upon DDB2 Binding Justifies the Different Repair Rates for Cyclobutane Pyrimidine Dimers.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.10.724147v1?rss=1">
<title>
<![CDATA[
Hearts may grow concentrically to balance ATP supply and demand and eccentrically to stabilize titin-based stress 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.10.724147v1?rss=1
</link>
<description><![CDATA[
Hearts change their wall thickness (concentric growth) and chamber size (eccentric growth) as they adapt to circulatory demands and the intrinsic function of their contractile cells. Factors associated with wall thickening include variants of sarcomeric proteins that enhance contractility, mitochondrial dysfunction, and hypertension. Chambers can dilate due to many factors including sarcomeric variants that depress contractility and aortic and / or mitral valve insufficiency. Despite intensive study, the mechanisms that regulate cardiac growth remain unclear. It is also uncertain whether inherited variants induce growth via the same mechanisms as more common clinical pathologies, such as hypertension. Here we show that computer simulations of a beating left ventricle reproduce both variant and non-variant-related growth patterns when myocytes grow concentrically to regulate intracellular ATP concentration and eccentrically to maintain titin-based intracellular stress. The simulations support the hypothesis that cardiac growth reflects homeostatic feedback through three interacting systems whereby myocytes add or remove mitochondria and sarcomeres (1) in parallel to match ATP generation to myocardial energy demand, and (2) in series to regulate passive tension, while (3) the autonomic nervous system regulates cardiac power, and thus myocardial ATPase, via baroreflex control. The new framework provides a mechanistic basis for the patterns of eccentric and concentric growth induced by a wide range of clinically-relevant conditions and could facilitate in silico testing of potential therapies for cardiac disease.
]]></description>
<dc:creator><![CDATA[ Kotter, J. R., Leung, S. W., Kampourakis, T., Lee, L.-C., Wenk, J., Moulton, M., Tanner, B. C. W., Campbell, S., Yengo, C. M., McDonald, K. S., Stelzer, J., Campbell, K. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.10.724147</dc:identifier>
<dc:title><![CDATA[Hearts may grow concentrically to balance ATP supply and demand and eccentrically to stabilize titin-based stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.10.724047v1?rss=1">
<title>
<![CDATA[
Beyond Redfield: Thermodynamic Bounds and Non-Perturbative Quantum Dynamics in Tubulin Networks 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.10.724047v1?rss=1
</link>
<description><![CDATA[
Quantum effects in biology are unavoidable at the molecular scale; the unresolved question is whether they can remain functionally relevant across the timescale gap between femtosecond molecular dynamics and microsecond-to-millisecond biological function. Here we formalize this mismatch as an equilibrium-to-functionality gap and use tubulin as a stringent open-system test case. We combine secular Lindblad, Redfield, and hierarchical equations of motion (HEOM) treatments to quantify decoherence, non-perturbative relaxation, and the physical amplification required for functional relevance. Equilibrium dephasing yields a conservative T2* {approx} 39 fs at 310 K, with a generic protein-bath baseline of {approx} 13 fs. A completed 30 ps HEOM trajectory for the full 1JFF tryptophan network shows distributed non-Markovian relaxation, with terminal purity Pur = 0.210 and stretched-exponential exponent beta_KWW {approx} 0.44, confirming that Redfield is useful as a short-time perturbative comparator but not quantitatively interchangeable with HEOM in this intermediate-coupling regime. We introduce a coherence-utility criterion U = K tau_coh / tau_func, separating required amplification from empirically bounded gain. A thermodynamic uncertainty relation closure shows that neural-scale cascade amplification would require P_min ~ 10^-7 W, about five orders of magnitude above the local microtubule GTP budget. Frohlich pumping is found to be linewidth-gated rather than generically micron-scale; ordered-water cavity QED and geometric subradiance remain experimentally testable but severely constrained candidates. The result is not a model of consciousness, but a reproducible physical benchmark framework for evaluating biological quantum-coherence claims under explicit open-system, energetic, and experimental constraints. Six falsifiable experimental programmes are prioritized, and the full computational framework is released with a validation ledger, cryptographic audit trail, and living supplementary material.
]]></description>
<dc:creator><![CDATA[ Firmenich, F., Firmenich, P., Firmenich, L. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.10.724047</dc:identifier>
<dc:title><![CDATA[Beyond Redfield: Thermodynamic Bounds and Non-Perturbative Quantum Dynamics in Tubulin Networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.09.724020v1?rss=1">
<title>
<![CDATA[
Proton-coupled alternating access in a versatile Spns drug efflux pump from Mycobacterium smegmatis 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.09.724020v1?rss=1
</link>
<description><![CDATA[
Spns transporters are major facilitator superfamily proteins that regulate lipid transport, lysosomal homeostasis, immunity and disease, yet how protonation enables their chemically diverse transport functions remains unclear. Here, we combine double electron-electron resonance spectroscopy in lipid nanodiscs with DEER- and AlphaFold-guided modeling to define the conformational landscape of the Mycobacterium smegmatis Spns homolog MsSpns. Protonation shifts MsSpns toward an inward-facing state, whereas deprotonation favors a broader outward-facing ensemble through coordinated rearrangements of the intracellular and extracellular gates. These transitions are governed by membrane-embedded protonation switches and proton-sensing networks on both sides of the membrane, while the substrate-binding cavity exhibits distinct proton sensitivity and weaker cooperativity. Hydrophilic cationic substrates, including capreomycin and ethidium bromide, stabilize the outward-facing state, consistent with efflux antiport, whereas lipophilic compounds, including rifampicin, epicholesterol and certain phospholipids, favor the inward-facing state, suggesting uptake or allosteric stabilization. Thus, conserved proton-coupling elements can drive substrate transport in opposite directions, revealing the mechanistic versatility of the Spns fold with therapeutic potential.
]]></description>
<dc:creator><![CDATA[ Gies, S., Jagessar, K. L., Wu, T., Miller, I., Dastvan, K., Dastvan, R. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.09.724020</dc:identifier>
<dc:title><![CDATA[Proton-coupled alternating access in a versatile Spns drug efflux pump from Mycobacterium smegmatis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.09.724055v1?rss=1">
<title>
<![CDATA[
Sustainable Technology for the Fabrication of Liposomal Phases 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.09.724055v1?rss=1
</link>
<description><![CDATA[
Liposomes are self-assembled lipid vesicles capable of encapsulating both hydrophilic and hydrophobic therapeutics, making them versatile platforms in drug delivery and biomedical technology. In this study, the limitations of the classical thin-film hydration method were critically evaluated, and a sustainable, systematically optimized strategy was established for generating defined liposomal lamellar phases. Hydration conditions were optimized, and 4 mL of buffer per 10 mg of lipid was determined to be optimal for effective rehydration and improved statistical reliability of vesicle measurements. A refined probe-sonication protocol (20% amplitude, 5 s ON/55 s OFF pulse) enabled controlled transformation of multivesicular vesicles into stable multilamellar and unilamellar vesicles at net ON-times of 90 s and 185 s, respectively, without overheating or contamination. In addition, a Python-based machine-learning tool was developed for vesicle size characterization. Collectively, these optimizations provided a reproducible and sustainable framework for preparing liposomes across different lamellar phases.
]]></description>
<dc:creator><![CDATA[ Polley, A., Ravikumar, A., Shanmugam, S. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.09.724055</dc:identifier>
<dc:title><![CDATA[Sustainable Technology for the Fabrication of Liposomal Phases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.09.721219v1?rss=1">
<title>
<![CDATA[
Benthic diatoms navigate shear flows via hydrodynamic rolling and active gliding 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.09.721219v1?rss=1
</link>
<description><![CDATA[
Navigating fluid flow is a fundamental challenge for microbial life across diverse aquatic environments. While rheotaxis in swimming microorganisms has been extensively studied, it remains unresolved whether near-bed shear merely perturbs gliding motility or instead provides directional cues for active navigation on surfaces. Here we show that the benthic diatom Navicula cryptocephala utilises a purely mechanical strategy to achieve downstream rheotaxis and anisotropic spreading on submerged surfaces. Single-cell ellipsoidal tracking reveals a direction-dependent angular response that reorients gliding cells towards the downstream direction. Using interference reflection microscopy, we further reveal that shear induces rolling of obliquely gliding cells, laterally shifting the cell-substrate contact site. This shift renders raphe-based propulsion non-collinear with substrate friction, generating a downstream-restoring yaw torque. Crucially, our results rule out alternative explanations based on longitudinal shifts of the raphe contact site or direct hydrodynamic yaw torque. A minimal stochastic model confirms that this mechanical reorientation alone is sufficient to reproduce the observed drift and diffusion patterns, without invoking either orientation-dependent switching between motility states or orientation-dependent dwell times of those states. Our findings uncover a mechanism by which ambient shear is converted into directional guidance for active surface motility, providing new insights into microbial transport, retention, and resilience on submerged surfaces.
]]></description>
<dc:creator><![CDATA[ Wang, B., Ding, S., Jiang, W., Guo, X., Han, R., Zeng, L., Wang, J., Pedley, T. J. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.09.721219</dc:identifier>
<dc:title><![CDATA[Benthic diatoms navigate shear flows via hydrodynamic rolling and active gliding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.09.723774v1?rss=1">
<title>
<![CDATA[
Dual curvature sensing governs cell orientation and curvotaxis 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.09.723774v1?rss=1
</link>
<description><![CDATA[
Cells lying in a curved environment can respond to the surface curvature by reorienting their shape. However, whether cells respond to the mean curvature and/or the Gaussian curvature remains largely unexplored. Here, inspired by experimental observations of how ovarian theca cells (TCs) orient themselves on substrates with different curvatures, we propose a theoretical framework for active nematic layers on curved surfaces. In this model, we assume that the nematic directors of the cells respond to both the mean curvature and the Gaussian curvature of the underlying substrate surface. Our theory predicts specific cell orientation patterns on hemicylindrical, hourglass- and dome-like substrates, consistent with experimental observations. In addition, by incorporating curvature-induced active traction, our model successfully recapitulates the experimental observation of TC accumulation at convex regions of hemicylindrical substrates as well as saddle-shaped regions of more complex geometries. Overall, our work reveals the unexpected role of cell curvature sensing in driving collective migration and pattern formation on various substrate curvature.
]]></description>
<dc:creator><![CDATA[ Chan, C., Lin, S.-Z., Tomida, K., Ng, B. H., Lee, C. H., Lee, J. S., Zhao, Z., Eliza, F. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.09.723774</dc:identifier>
<dc:title><![CDATA[Dual curvature sensing governs cell orientation and curvotaxis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723928v1?rss=1">
<title>
<![CDATA[
AI-discovered protein fragments as generalizable regulators of biomolecular condensates 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723928v1?rss=1
</link>
<description><![CDATA[
Biomolecular condensates are a major driver of cellular organization; however, we lack a predictable and systematic approach to modulate the multivalent interactions underlying their formation. Here, we demonstrate that the AI-driven FragFold method enables robust and generalizable design of protein fragments to control biomolecular condensate formation. We apply this approach across diverse proteins: G3BP1, SARS-CoV-2 nucleocapsid, TDP-43, and focal adhesion kinase (FAK). Computationally screening 2,235 fragments, we selected 18 candidates for further investigation. Overall, we attain a 50% success rate (9/18 designs) in discovering condensate-controlling protein fragments, experimentally testing just 3-5 candidates per protein. For each condensate-forming protein, the success rate is at least 40%. Furthermore, FragFold-predicted fragment binding modes align with their condensate-inhibitory or -enhancing activities, revealing both known and newly identified interactions underlying condensate formation. In FAK, a condensate-inhibitory fragment uncovered a domain interaction required for phase separation, and mutational analysis validated its importance. Notably, this inhibitory fragment also suppresses FAK condensate formation in living mammalian cells. Together, these results establish AI-guided protein fragment discovery as a generalizable strategy to dissect and control the molecular interactions that govern biomolecular condensates.
]]></description>
<dc:creator><![CDATA[ Savinov, A., Sadasivan, J., White, K. J., Rubien, J. D., Li, G.-W., Case, L. B. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723928</dc:identifier>
<dc:title><![CDATA[AI-discovered protein fragments as generalizable regulators of biomolecular condensates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723821v1?rss=1">
<title>
<![CDATA[
Repulsion-Driven Layering in Polymer-Assisted Condensation 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723821v1?rss=1
</link>
<description><![CDATA[
The hierarchical organization of multiphase biomolecular condensates into core-shell architectures is a fundamental problem in soft matter and biophysics. While classical explanations rely on hierarchies of interfacial tension ({gamma}) between coexisting liquids, the ultralow tensions of condensates (0.1-1 {micro}N/m) render such hierarchies potentially fragile. We introduce a robust assembly principle based on Polymer-Assisted Condensation (PAC), in which a single polymer species dictates the entire structure. The polymer nucleates a dense core by recruiting a condensation-incompetent protein (P1). A second incompetent protein (P2), which is repelled or otherwise thermodynamically disfavored from entering the polymer-rich core, is nonetheless recruited to the interface by weak attraction to P1, forming a stable shell. This effective repulsion-driven layering operates across a wide parameter space without requiring{gamma} asymmetries and yields a robust structure that is impervious to concentration fluctuations and environmental perturbations. Phase-field modeling and molecular simulations establish this mechanism and capture key features of nucleolar organization. Our work reveals a general physical pathway for encoding spatial order in soft, multicomponent fluids.
]]></description>
<dc:creator><![CDATA[ Majee, A., Merlitz, H., Schiessel, H., Sommer, J.-U. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723821</dc:identifier>
<dc:title><![CDATA[Repulsion-Driven Layering in Polymer-Assisted Condensation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723778v1?rss=1">
<title>
<![CDATA[
Optical single-channel recording of CRAC channels with HaloTag and a Ca2+-sensitive ligand 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723778v1?rss=1
</link>
<description><![CDATA[
Following ER Ca2+ depletion, Ca2+ release-activated Ca2+ (CRAC) channels are activated by STIM1 at ER-plasma membrane junctions. The restricted localization and low conductance of the CRAC channel (<40 fS) precludes single-channel recordings, limiting studies of CRAC channel gating. Here we describe an optical approach to characterize the gating of HaloTag-fused Orai1 channels labeled with JF646-BAPTA, a Ca2+-sensitive fluorescent dye. While Ca2+ influx through single channels generates fluorescence fluctuations, identifying true gating events is complicated by stochastic transitions of JF646-BAPTA to a non-fluorescent state. To overcome this, we combine TIRF microscopy with whole-cell voltage clamp to control the driving force for Ca2+ entry. We show the open channel intensity at -100 mV reflects Ca2+ saturation of the dyes on each channel, while the closed-channel intensity is defined by the fluorescence at +30 mV, where influx is absent. True gating events can be identified from transitions between the open- and closed-channel levels, distinguishing them from transitions to a non-fluorescent state. We describe the gating behavior of CRAC channels activated by STIM1 after store depletion. Dwell time distributions indicate at least two open and closed states with durations of 0.1 to several seconds, with most channels having an open probability of [&ge;]0.7. We also detect  silent channels that colocalize with STIM1 but show no activity over tens of seconds, a population that would be undetectable by whole-cell electrophysiology alone. This method offers an approach to explore CRAC channel gating mechanisms and may be applicable to other Ca2+- permeable channels not amenable to patch-clamp techniques.
]]></description>
<dc:creator><![CDATA[ Dhillon, H., Lewis, R. S. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723778</dc:identifier>
<dc:title><![CDATA[Optical single-channel recording of CRAC channels with HaloTag and a Ca2+-sensitive ligand]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723761v1?rss=1">
<title>
<![CDATA[
Using iPALM to determine protein organisation in cardiac muscle Z-discs 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723761v1?rss=1
</link>
<description><![CDATA[
Sarcomeres, the basic repeating unit of striated muscle, are joined together by crosslinked actin filaments found at the boundaries of muscle sarcomeres, termed Z-discs. Z-discs play a key role in cardiac signalling and disease, however, the arrangement and function of many of the proteins present in the Z-disc remain to be understood. Here, we determined the organisation of 3 key proteins, ZASP, [a]-Actinin-2 and the Z1Z2 epitope of titin, located within the Z-disc. We fluorescently labelled these proteins in cardiac myofibrils using Adhirons specific to each protein and used interferometric photoactivated localization microscopy (iPALM) to obtain the 3D position of these proteins to a high precision (<10nm in x,y,z). We then used PERPL (Pattern Extraction from Relative Positions of Localisations) to analyse patterns in the relative positions of the proteins and reveal their underlying organisation. This analysis revealed that ZASP and [a]-Actinin-2 have a similar repeating organisation, but that the organisation of Z1Z2 is different.
]]></description>
<dc:creator><![CDATA[ Umney, O., Curd, A. P., Martin, H., Lewis, T., Tang, A. A.-S., Balusubramanian, H., Khuon, S., Aaron, J., Peckham, M. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723761</dc:identifier>
<dc:title><![CDATA[Using iPALM to determine protein organisation in cardiac muscle Z-discs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723777v1?rss=1">
<title>
<![CDATA[
Yoda molecules agonize PIEZO2 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723777v1?rss=1
</link>
<description><![CDATA[
PIEZO proteins (PIEZO1 and PIEZO2) are essential mechanosensitive channels. PIEZO1 is thought to be selectively activated by Yoda molecules (Yoda1 and Yoda2). Although a structural framework for PIEZO1 activation by Yoda1 exists, a molecular mechanism underlying this selective activation is lacking. Here, using electrophysiology and calcium imaging, we show that Yoda1 increases PIEZO2 open probability and stretch sensitivity as efficaciously as PIEZO1 but elicits weaker PIEZO2-dependent calcium entry, rationalizing why its effect on PIEZO2 has been overlooked. Both Yoda1 and its more potent Yoda2 analog slow down inactivation of PIEZO2 currents with potency similar to PIEZO1 but with lower efficacy. Using mutagenesis and molecular dynamics simulations, we further show that Yoda2s benzoic acid group forms a transient salt bridge with a conserved arginine in the Yoda binding site, providing a molecular basis for Yoda2s increased potency. Our study cautions a reevaluation of studies using these molecules to untangle biological functions mediated by PIEZO channels.
]]></description>
<dc:creator><![CDATA[ Wijerathne, T. D., Chandrasekharan, A., Bhatt, A., Luo, Y. L., Lacroix, J. J. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723777</dc:identifier>
<dc:title><![CDATA[Yoda molecules agonize PIEZO2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723694v1?rss=1">
<title>
<![CDATA[
Non-covalent reversibly photoconvertible fluorescent tags for wash-free protein labeling 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723694v1?rss=1
</link>
<description><![CDATA[
Reversibly photoswitchable fluorophores are widely used in advanced bioimaging but their design remains demanding. Here, we introduce a new series spanning the whole visible range, which results from combining a large set of fluorogens with the FAST protein scaffold. We first demonstrate that these well-established labeling fluorescent protein tags turn into negative reversible photoswitchers upon decreasing the fluorogen concentration and increasing light intensity. We then show that using not anymore one but two fluorogens adds new responses to illumination. Thus, we obtain positive reversible photoswitchers, that increase their brightness under illumination. We also generate a palette of non-covalent reversibly photoconvertible fluorescent proteins changing their fluorescence color upon illumination, a reversible behavior that still remains absent in regular fluorescent proteins. This light-induced color change opens the possibility to discriminate six spectrally similar FAST variants in live cells upon demonstrating the superiority of using multiple spectral channels for exploiting the time dependence of the fluorescence response to illumination.
]]></description>
<dc:creator><![CDATA[ Mandal, M., Shpinov, Y., Lahlou, A., Pham, F., El Hajji, L., Coghill, I., Laureau, E., Plamont, M.-A., Perez, F., Le Saux, T., Aujard, I., Gautier, A., JULLIEN, L. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723694</dc:identifier>
<dc:title><![CDATA[Non-covalent reversibly photoconvertible fluorescent tags for wash-free protein labeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
