Biology subjects

Zhang, G.

Publications and source records attributed to Zhang, G..

At least 109 records · Page 6Linked to original sources

bioRxiv2025

M-DeepAssembly2: A Web Server for Predicting Multiple Conformations of Multi-domain Proteins Using Deep Learning

The biological functions of proteins often depend on dynamic transitions between multiple conformational states, and for multi-domain proteins, inter-domain dynamics play a critical role in their biological functions. Despite significant advances in static structure prediction, accurately modeling these multiple conformations remains a major challenge. Here, we present M-DeepAssembly2, a web server designed for modeling multiple conformations of multi-domain proteins using deep learning-predicted inter-domain and intra-domain constraints. The server operates in three stages. Firstly, evolutionary, physicochemical, and geometric features are extracted from the input and fed into AlphaFlex, a flexible residue prediction network. This process generates multiple distance maps that effectively capture both inter-domain and intra-domain interactions. Secondly, supplementary distance maps that characterize inter-domain interactions are generated using the DeepAssembly method. Finally, all distance maps are integrated as constraints in a multi-objective protein optimization to generate multiple conformational states of the protein. Compared to its predecessor, M-DeepAssembly2 leverages a flexible residue prediction network that directionally decouples coevolutionary information to generate heterogeneous distance maps. Benchmark results demonstrate that this strategy enables accurate prediction of protein multiple conformations and significantly improves the precision of static multi-domain protein structure prediction.

bioinformatics↗

bioRxiv2025

ProSiteHunter: A unified framework for sequence-based prediction of protein-nucleic acid and protein-protein binding sites

Accurate prediction of protein binding sites is essential for elucidating protein function, understanding molecular interaction mechanisms, and facilitating drug design. However, existing sequence-based approaches are often designed for specific binding-site types and therefore lack generality, whereas structure-based methods typically rely on high-quality structural models, limiting their applicability. Here, we introduce ProSiteHunter, a unified sequence-based framework for protein binding-site prediction, which integrates a fine-tuned protein language model (SiteT5) with a multi-source feature-fusion network that incorporates evolutionary, geometric, and statistical features, while employing bidirectional semantics, local associations, and global dependencies for comprehensive binding-site characterization. The method was systematically evaluated on diverse binding sites prediction tasks, where ProSiteHunter achieved a 39.1% average improvement in PRAUC for protein-DNA/RNA/protein tasks and a 7.4% PRAUC enhancement on the particularly challenging antibody-antigen task over state-of-the-art methods. Moreover, ProSiteHunter is capable of identifying local flexible sites that complement AlphaFold3 predictions and improving the accuracy of antibody-antigen interaction prediction. These results highlight ProSiteHunter as an efficient and unified approach for accurate and robust prediction of diverse protein binding sites.

bioinformatics↗

bioRxiv2025

Migration patterns and hybridization within the Asian stonechat complex in response to a major geographical barrier

Long-distance avian migration is thought to be under strong natural selection. Facing geographical barriers, migrants display various patterns considered to be adaptive. For example, they may detour along either side around the barrier or cross it, requiring specialized behavioral adaptations. Variations within closely related taxa are excellent sources for understanding the evolutionary background of migration and how barriers are shaping migration routes. In Asia, some species are assumed to have a migratory divide in response to the major geographical barrier, the Qinghai-Tibet Plateau (QTP), including the stonechat taxa (Siberian Stonechat Saxicola maurus maurus and Amur Stonechat S. stejnegeri). As they detour along either side of the QTP, these taxa are believed to disfavor a crossing over the highland. However, the more southernly distributed Tibetan Stonechat (S. m. przewalskii) breeds on the QTP, suggesting adaptation to high elevation. To investigate migration patterns and the potentially associated genetic differences, we studied migration routes and population genetics of four populations around the assumed migratory divide in Russia and Mongolia, and of one from the QTP in China. Our results confirmed the existence of a migratory divide between maurus and stejnegeri, albeit with extensive hybridization. We observed both the hypothesized western and eastern routes, but also found individuals employing intermediate routes crossing the QTP, of which two-thirds were clear hybrids. Meanwhile, przewalskii followed a highland-crossing route and was genetically differentiated from maurus and stejnegeri. The diverse migration routes among Asian stonechats show differential responses towards the geographical barrier. The intermediate route may be associated with hybridization, and its conditional viability may facilitate gene flow between maurus and stejnegeri. The Asian stonechat complex thus offers great opportunities for novel research of the genetics and evolution of migration. The specific evolutionary background associated with inhabiting and crossing the QTP can offer new perspectives in this field. Teaser textMigratory divides can arise in birds because alternative routes around migratory barriers would select for behaviors to restrict hybridization. Hybrids of parental types that employ alternative routes are hypothesized to embark on intermediate routes that would expose them to suboptimal conditions, resulting in post-zygotic reproductive isolation. However, this hypothesis is challenged when a sister taxon actually breeds on the geographical barrier. This is the case in the Asian stonechat complex that breeds near or on the Qinghai-Tibet Plateau (QTP), the roof of the world. We demonstrated a migratory divide in central Siberia to Mongolia for race maurus and stejnegeri, yet showed also evidence for extensive hybridization. Hybrids migrated along a newly discovered intermediate route, seemingly viable and overlaps with the migration trajectory of race przewalskii over the eastern part of the QTP. The Asian migratory divide relative to the QTP thus provided new insights to the evolution of landbird migration.

evolutionary biology↗

bioRxiv2025

BubR1 TPR domain supports mitotic checkpoint by promoting MCC formation and MCC-APC/C interaction

BubR1 is the key component of the mitotic checkpoint, a surveillance mechanism ensures accurate chromosome segregation by facilitating the assembly of the mitotic checkpoint complex and promoting its binding to the anaphase-promoting complex/cyclosome. Although BubR1s role in SAC signaling has been extensively investigated, the function of its N-terminal tetratricopeptide repeat (TPR) domain remains poorly understood. In this study, we first established the essential role of the BubR1 TPR domain in SAC signaling. Guided by the resolved cryo-EM structure of the MCC-APC/C complex, we identified and characterized several interactions involving this domain with Mad2, Cdc20APC/C, Apc2. Furthermore, we discovered an intramolecular interaction between the TPR domain and downstream residues of BubR1, which appears to organize a structure resembling a "lasso" that incorporates four Cdc20APC/C-binding elements, thereby enhancing engagement with Cdc20 in the APC/C. Functional and biochemical analyses demonstrated that these interactions collectively promote MCC assembly and MCC-APC/C binding, enabling rapid SAC activation in response to microtubule-kinetochore attachment defects.

cell biology↗

bioRxiv2025

Fragile nucleosomes are essential for RNA Polymerase II to transcribe in eukaryotes

Nucleosomes are barriers to RNA Polymerase II (Pol II) transcribing along gene bodies in eukaryotes. We found that a fragile "tailless nucleosome" could be generated to resolve the Pol II pausing caused by the +1 nucleosome in metazoans. We hypothesized that "pan-acetylated nucleosomes" along gene bodies are fragile and are generated to resolve the resistance of nucleosomes to the elongation of Pol II. To confirm this hypothesis, we systematically analyzed pan-acetylation and pan-phosphorylation of the entire human genome in HEK293T cells. Herein, we show that both pan-acetylation and pan-phosphorylation of histones in nucleosomes along gene bodies are coupled with the activities of Pol II. Our results show that there are four major types of fragile nucleosomes required for Pol II to transcribe along gene bodies: nucleosomes without tails, nucleosomes with dominant acetylation, nucleosomes with dominant phosphorylation, and nucleosomes with both acetylation and phosphorylation.

genetics↗

bioRxiv2025

Quinolinic acid metabolism may mitigate AKI to CKD transition

The transition from acute kidney injury (AKI) to chronic kidney disease (CKD) remains a significant clinical problem with unclear underlying mechanisms. Emerging evidence suggests that alterations in tryptophan metabolism, particularly in the production of downstream metabolites such as quinolinic acid (QA), play a role in renal pathophysiology. QA is a NAD biosynthesis intermediate metabolized by the enzyme quinolinate phosphoribosyltransferase (QPRT). In this study, we investigated the role of QA in the AKI-to-CKD transition using experimental mouse models and clinical observations and leveraging multiple omics approaches. Systematic metabolomic profiling identified endogenous QA as one of the most significantly elevated metabolites following folic acid-(FA) induced injury. Exogenous QA exacerbated FA-induced kidney dysfunction. Conversely, aged mice deficient in QPRT showed worsened expression of kidney fibrosis markers even in absence of kidney injury, while younger littermates exhibited worsened induced kidney injury. Mice lacking QA-producing enzymes resisted experimental AKI and AKI-to-CKD progression. Multimodal spatial metabolomics analysis of human AKI kidney biopsies revealed QA accumulation in regions of inflammatory infiltration. Finally, children with CKD exhibited higher urinary QA levels compared to healthy controls. These findings underscore QA as a potential mediator of kidney injury and a therapeutic target for preventing the progression from AKI to CKD. One Sentence SummaryQuinolinic acid promotes kidney damage and fibrosis, suggesting it as a contributor of AKI-to-CKD progression and a potential therapeutic target.

pathology↗

bioRxiv2025

Application of MT3 in the Development and Immunogenicity Evaluation of GnRH Vaccines

Research on immune castration vaccines is of great significance for animal management. In this study, five copies of GnRH were linked to two MT3s by a flexible linker, GGGGS, to construct a recombinant plasmid, pET30a-GnRH5-MT3, which was expressed and purified using E. coli BL21(DE3) cells. The recombinant protein was best expressed under the induction conditions of 25 with 0.25 mM IPTG, with a purity of up to 80% after purification. It was mixed with the external adjuvant ISA 206 and immunized mice by subcutaneous injection. Serum levels of GnRH-specific Antibody, T, FSH and LH were detected by ELISA, and testicular tissue morphology was observed by HE staining. The results showed that GnRH5-MT3 recombinant protein combined with external adjuvant could efficiently induce GnRH-specific antibody production in male mice (P < 0.01-0.001) and significantly reduce the Serum LH, FSH and T concentrations (P < 0.01-0.001). The immunized mice had sparse cells in the seminiferous tubules of the testis, significantly reduced cells at various developmental stages, and almost no mature spermatozoa. This study provides experimental basis for further application of the vaccine in animal immune castration.

immunology↗

bioRxiv2025

ATF4 alleviates doxorubicin-induced cardiomyopathy through H2S-mediated antioxidation

BackgroundDoxorubicin (DOX) remains a cornerstone in the treatment of various cancers. However, its clinical utilization is significantly hampered by dose-dependent cardiotoxicity. The generation of reactive oxygen species (ROS) constitutes the central component of the pathogenesis of DOX-induced cardiotoxicity. Activating transcription factor 4 (ATF4) has been demonstrated to exert a cardioprotective effect and augment cardiac antioxidative capacity in settings of heart failure. However, the role of ATF4 in DOX-induced cardiomyopathy (DIC) remains unknown. MethodsTo explore the role of ATF4 in DOX-induced cardiomyopathy, cardiac-specific ATF4 conditional heterozygous mice and AAV9 mediated ATF4 overexpression mouse models were utilized. Cardiac function was assessed by echocardiography. The upstream regulator and downstream mediator of ATF4 were evaluated using RNA-seq analysis and further verified using ChIP assay and luciferase reporter assay. ResultsWe found a substantial decrease in ATF4 expression levels in the heart of DIC mice. ATF4+/- mice exhibited a higher degree of susceptibility to DOX-induced cardiotoxicity in comparison with ATF4flox/flox mice, as evidenced by the manifestation of more severe cardiac dysfunction and a significantly earlier mortality rate. In contrast, cardiacc-specific overexpression ATF4 by AAV9 confers robust cardioprotection against DOX-induced cardiomyopathy. Mechanistically, we identified the upstream regulator of ATF4 as KLF16, which was significantly suppressed during DOX treatment. Further, the decrease of ATF4 led to a reduction in cystathionine {gamma}-lyase (CSE) transcription and hydrogen sulfide (H2S) production in the context of DOX-induced cardiotoxicity. ChIP and luciferase reporter assays revealed that ATF4 functioned as the transcription factor of the CSE gene, which is a key enzyme in the synthesis of H2S to counteract oxidative stress. Consistently, ROS scavengers or H2S donors was shown to mitigate the consequences of ATF4 deficiency. In contrast, the ectopic expression of ATF4 mitigated oxidative stress and apoptosis in DOX-induced cardiotoxicity, both in vivo and in vitro. ConclusionsOur study revealed a novel function of ATF4 in counteracting oxidative stress in DOX cardiotoxicity by promoting the transcription of CSE. ATF4 may represent a promising therapeutic target for the treatment of DOX-induced cardiomyopathy.

pathology↗

bioRxiv2025

Comparative snRNAseq study of C9orf72, SOD1, and sALS spinal cord

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease characterized by the loss of motor neurons, yet the cell-type specific molecular alterations within the spinal cords are not well characterized. In this study, we conducted deep molecular profiling of spinal cord tissues donated by people living with sporadic, C9orf72, and SOD1-ALS using single-nucleus RNA sequencing (snRNAseq). We observed numerous distinct gene expression patterns and enriched pathways among ALS types. However, when focusing on common features, we identified activation of stress-response and inflammatory pathways in specific microglia subtypes, as well as disrupted vesicle transport and synaptic function in a ventral inhibitory neuronal subtype. Notably, CPLX3, a SNARE regulator, was uniquely expressed in alpha-motor neurons and was commonly downregulated across all ALS types. While this study uncovers the molecular heterogeneity underlying ALS, it also highlights shared pathways within specific cell types, especially in the ventral inhibitory neurons that have been less explored in ALS research.

neuroscience↗

bioRxiv2025

Structural insights and engineering of deep-sea halophilic PET hydrolytic enzymes

Pervasive use of polyethylene terephthalate (PET) poses tremendous challenges for global waste management and environmental sustainability, fueling growing interests in enzymatic degradation as an eco-friendly solution. While PET hydrolases hold significant promise, their industrial deployment is hindered by insufficient performance, particularly under high-salinity conditions raised from high substrate loads. Building on our previous discovery of three deep-sea PET hydrolases (dsPETase01, dsPETase05 and dsPETase06) with exceptional halophilicity and PET-degrading activity, we here present their three-dimensional structures and mechanistic characterization. Structural comparison, site-directed mutagenesis, and domain swapping reveal key structural features and the essential role of C-terminal acidic residues in salt tolerance. Integrating semi-rational protein design, Transformer-based modeling, and disulfide bond engineering, we synergistically enhance the thermostability and catalytic efficiency of dsPETase05. These findings elucidate the unique structural features and salt adaptation mechanisms of deep-sea halophilic PET hydrolases, and inform the future engineering of biocatalysts for application in harsh industrial environments.

molecular biology↗

bioRxiv2025

IRE1α-XBP1s axis is required for BCG-induced trained immunity against viral infection by inhibiting AHR signaling

Although Bacille Calmette-Guerin (BCG) has been proved to be suitable in inducing trained immunity, mechanism-related issues have not yet been fully resolved. In this study, we found that in vitro BCG training protects both BCG harboring and bystander macrophages against DNA virus and RNA virus infection and demonstrated that BCG harboring cells protect bystanders partly by type I interferon (IFN). We determined aryl hydrocarbon receptor (AHR) as a negative regulator in BCG-induced trained immunity on defense against viral infection in vitro and in vivo and blockade of type I IFN signaling partially impaired the enhanced antiviral effect of AHR deficiency in BCG training. Furthermore, we revealed IRE1-XBP1s signaling as a critical pathway in BCG-induced trained immunity against viral infection in vitro and in mice. Mechanistically, we demonstrated that IRE1-XBP1s pathway influenced BCG-induced trained immunity by limiting AHR mRNA expression and the translocation of AHR protein into the nucleus.

immunology↗

bioRxiv2025

GFFx: A Rust-based suite of utilities for ultra-fast genomic feature extraction

Genome annotations are becoming increasingly comprehensive due to the discovery of diverse regulatory elements and transcript variants. However, this improvement in annotation resolution poses major challenges for efficient querying, especially across large genomes and pangenomes. Existing tools often exhibit performance bottlenecks when handling large-scale genome annotation files, particularly for region-based queries and hierarchical model extraction. Here, we present GFFx, a Rust-based toolkit for ultra-fast and scalable genome annotation access. GFFx introduces a compact, model-aware indexing system inspired by binning strategies and leverages Rusts strengths in execution speed, memory safety, and multithreading. It supports both feature- and region-based extraction with significant improvements in runtime and scalability over existing tools. Distributed via Cargo, GFFx provides a cross-platform command-line interface and a reusable library with a clean API, enabling seamless integration into custom pipelines. Benchmark results demonstrate that GFFx offers substantial speedups and makes a practical, extensible solution for genome annotation workflows.

bioinformatics↗

bioRxiv2025

Reengineering the antigen optimization process for superior neoantigen vaccine design

Identifying effective neoantigen sequences is essential for enhancing anti-tumor immunity. However, the vast sequence space (>109 possible peptides) and limited accuracy of existing immunogenicity predictors hinder efficient vaccine design for patient-specific human leukocyte antigens (HLAs). We present AlphaVacc, a deep reinforcement learning framework that integrates Monte Carlo Tree Search with a Transformer-based network to optimize antigenic peptides. AlphaVacc outperforms previous generative models in binding-affinity prediction. Experimental validation of 12 AlphaVacc-generated variants of the BING-4 peptide confirmed that 11 showed increased HLA-A*02:01 binding and 7 elicited significant T cell responses. Further testing of 16 single-mutation peptides confirmed computational predictions for 15 candidates, exhibiting a remarkable success rate. AlphaVacc thus provides a powerful tool for designing neoantigen-based cancer vaccines and may accelerate personalized immunotherapies.

bioinformatics↗

bioRxiv2025

MViewEMA: Efficient Global Accuracy Estimation for Protein Complex Structural Models Using Multi-View Representation Learning

Estimation of model accuracy (EMA) is crucial for the reliable application of protein structure models in biological research. With the rapid advancement of protein structure prediction techniques and the explosive growth of predicted structural data, existing EMA methods struggle to balance computational efficiency with estimation performance. Here, we present MViewEMA, a single-model EMA method that leverages a multi-view representation learning framework to integrate residue-residue interaction features from micro-environment, meso-environment, and macro-environment levels for global accuracy assessment of protein complex models. Benchmark results demonstrate that MViewEMA outperforms state-of-the-art EMA methods in global accuracy assessment, achieving more than a 10-fold improvement in computational efficiency compared to our previous method, DeepUMQA3. This method enables efficient selection of high-quality protein complex models from large-scale structural datasets and achieved top performance in model selection tracks during the CASP16 blind test, demonstrating its potential to enhance the accuracy of complex structure prediction when integrated into modern frameworks such as AlphaFold-Multimer, AlphaFold3, and DiffDock-PP.

bioinformatics↗

bioRxiv2025

Type 1 Regulatory T cells induced by intestinal epithelial cells respond to food antigen

As the first-line barrier against food antigens, the intestinal epitheliums role in establishing peripheral immune tolerance to food remains completely unknown. Additionally, the specific types of regulatory T cells induced to maintain food tolerance--along with their antigen specificity--are still unclear. Here we found small intestinal FOXP3-IL-10+ Tr1 cells, but not Treg cells, can respond to food antigens presented by intestinal epithelial cells (IECs) in an antigen-specific manner. Through immunopeptidome analysis of MHCII-presented peptides, we demonstrate that IECs can present food-derived antigens. Importantly, these IEC-presented food antigens are exclusively recognized by the TCR repertoire of small intestinal Tr1 cells, not Treg cells. Furthermore, we show that T cells recognizing IEC-presented food antigens specifically differentiate into Tr1 cells in the siLP. These findings establish two key insights: first, that maintaining food tolerance represents a physiological function of Tr1 cells, and second, that IECs play a central role in maintaining food tolerance. Our work thus provides mechanistic understanding of how the intestinal epithelium contributes to immune tolerance through its antigen presentation capacity and subsequent Tr1 cell induction.

immunology↗

bioRxiv2025

Repulsive vs. Attractive Crowding Distinctly Regulate TDP-43 Condensates through Region-Specific Structural Dynamic

TAR DNA-binding protein 43 (TDP-43) is crucial for RNA processing and nucleocytoplasmic transport, and its pathological aggregation is a hallmark of neurodegenerative disorders such as amyotrophic lateral sclerosis and frontotemporal dementia. The intrinsically disordered, prion-like C-terminal domain (CTD) of TDP-43 drives its liquid-liquid phase separation (LLPS), a process fundamental to both normal cellular function and disease-associated aggregation. Using coarse-grained molecular simulations, we systematically explored how distinct macromolecular crowding environments, including repulsive (steric) and attractive (interaction-based) crowding conditions, influence the phase behavior and internal organization of TDP-43 CTD condensates. Our findings reveal that both repulsive and attractive crowders maintain strong correlations between single-chain compaction, dimerization propensity, and macroscopic phase separation, yet operate via distinct physical mechanisms: repulsive crowders drive condensation through volume-exclusion-mediated entropic stabilization, while attractive crowders modulate condensate formation via competitive enthalpic interactions. Spatial structural analysis identifies a robust region-specific internal architecture, with -helices enriched at the condensate core and aligned parallel to the interface, and intrinsically disordered regions (IDRs) preferentially localized near the condensate surface in orientations nearly perpendicular to the interface. Dynamically, -helical regions form strong yet transient interaction hubs, whereas IDRs establish weaker but more persistent contacts, thereby maintaining structural fluidity. Collectively, our results uncover a regulatory principle by which macromolecular crowding modulates TDP-43 condensation through distinct entropic and enthalpic contributions, offering key mechanistic insights into condensate formation and dysregulation relevant to neurodegenerative diseases.

biophysics↗

bioRxiv2025

Harmonic Amplitude-Modulated Singular Value Decomposition for Ultrafast Ultrasound Imaging of Gas Vesicles

Ultrafast nonlinear ultrasound imaging of gas vesicles (GV) contrast agents promises high-sensitivity biomolecular visualization with applications such as targeted molecular imaging of tumor markers or real-time tracking of gene expression. However, separating GV-specific signal from tissue remains challenging and requires the implementation of complex transmit schemes. In this work we introduce harmonic amplitude-modulated singular value decomposition (HAM-SVD), which synergizes pulse inversion (PI) with amplitude-modulated singular value decomposition (AM-SVD) to isolate GV-specific second-harmonic signals. In HAM-SVD, single-cycle plane waves at 9.6 MHz and five tilted angles (at a pulse repetition frequency of 2500 Hz) are transmitted under four duty cycles with alternating polarity. Beamformed IQ data are reshaped along a "space x pressure" matrix and decomposed via SVD; tissue background is cancelled by discarding the first and lowest singular modes, yielding an image comprised solely of pressure-dependent second harmonic signals. HAM-SVD sequence enables wide-field, ultrafast imaging without complex transmit sequences. Validation via simulations, in vitro phantoms, and in vivo rat lower limb experiments demonstrates HAM-SVDs outperformance compared to PI and AM-SVD. HAM-SVD is shown to achieve a 19.16 {+/-} 1.63 dB signal-to-background ratio (SBR) in vivo, surpassing PI (14.19 {+/-} 1.41 dB) and AM-SVD (15.79 {+/-} 1.38 dB). HAM-SVD overcomes limitations of conventional nonlinear techniques (e.g., depth restrictions, tissue clutter) by combining PIs harmonic sensitivity with AM-SVDs adaptive clutter filtering of tissue signals. This approach enhances molecular imaging specificity for GVs and holds potential for ultrasound localization microscopy of slow-flowing agents.

bioengineering↗

bioRxiv2025

AlphaFlex: Accuracy modeling of protein multiple conformations via predicted flexible residues

Understanding protein conformational dynamics is critical for elucidating disease mechanisms and facilitating structure-based drug design. While AlphaFold2-based methods have advanced static structure prediction, modeling multiple conformation states with both accuracy and efficiency remains challenging. We present AlphaFlex, an innovative framework that integrates flexible residue prediction with a directed masking strategy for multiple sequence alignments. By selectively relaxing co-evolutionary constraints in dynamic regions, AlphaFlex enables accurate prediction of biologically relevant states while preserving structural fidelity. Benchmarking on 69 apo-holo pairs from the CoDNaS dataset, including enzymes, binding proteins, and chaperones, reveals AlphaFlexs superior performance in both flexible residue identification and multiple conformation prediction. Comparative analysis against AF-Cluster, AF2-conformations, and AlphaFlow demonstrates AlphaFlexs significant advantage, achieving the highest success rate (42%) in accurately predicting apo/holo structures (TM-score >0.95 for both states) while consistently outperforming these methods in reproducing both apo and holo conformations. Notably, AlphaFlex maintains robust performance for membrane proteins, resolving inward- and outward-facing conformations with equal fidelity. Case studies further reveal its unique ability to sample physically plausible intermediate states along transition pathways. These results establish AlphaFlex as a pivotal computational tool for mapping protein conformational landscapes, with promising potential to accelerate structure-based therapeutic development.

bioinformatics↗