Protein Science最新文献_第5页

Total synthesis and structural characterization of a novel protein scaffold from the snail Biomphalaria glabrata. 一种新型光螺蛋白支架的合成及结构表征。

IF 5.6 3区生物学

Protein Science Pub Date : 2026-09-01 DOI: 10.1002/pro.70745

Oleg Melnyk, Stéphanie Caby, Armelle Vigouroux, Christine Demanche, Rémi Desmet, Magalie Sénéchal, Benoît Snella, Alexandra Mougel, Céline Boidin-Wichlacz, Aurélie Parmentier, Ugo Pasco, Sonia Cantel, Solange Moréra, Jérôme Vicogne

{"title":"Total synthesis and structural characterization of a novel protein scaffold from the snail Biomphalaria glabrata.","authors":"Oleg Melnyk, Stéphanie Caby, Armelle Vigouroux, Christine Demanche, Rémi Desmet, Magalie Sénéchal, Benoît Snella, Alexandra Mougel, Céline Boidin-Wichlacz, Aurélie Parmentier, Ugo Pasco, Sonia Cantel, Solange Moréra, Jérôme Vicogne","doi":"10.1002/pro.70745","DOIUrl":"10.1002/pro.70745","url":null,"abstract":"Disulfide-rich miniproteins constitute compact and highly stable scaffolds of growing interest for molecular and structural engineering. Schistosomins are ~80-residue proteins conserved across gastropods that form a long-standing orphan family whose structure and biological roles have remained unknown. Here, we report the total chemical synthesis and structural characterization of a schistosomin isoform from Biomphalaria glabrata, a medically relevant intermediate host of the parasite Schistosoma mansoni. Using state-of-the-art solid-phase peptide synthesis, chemoselective peptide ligation, and controlled oxidative folding, we obtained homogeneous, well-folded schistosomin suitable for biophysical and structural studies. High-resolution X-ray crystallography reveals a previously undescribed disulfide-rich fold defining a new class of miniprotein scaffold. Nano differential scanning fluorimetry and circular dichroism experiments demonstrate the remarkable thermal stability of this scaffold. Complementary in silico analyses suggest that the two naturally occurring isoforms, which differ by a single residue, exhibit highly similar structural and dynamic properties. Finally, transcript and protein analyses across snail tissues provide the first spatial expression map of schistosomin in a medically relevant mollusk. Together, this work establishes schistosomin as a novel and robust miniprotein scaffold and provides a structural and biological framework for exploring its function and potential applications.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70745"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13440164/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148674038","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Investigating enzyme function by geometric matching of catalytic motifs. 通过催化基序的几何匹配研究酶的功能。

IF 5.6 3区生物学

Protein Science Pub Date : 2026-09-01 DOI: 10.1002/pro.70744

Raymund E Hackett, Ioannis G Riziotis, Martin Larralde, António J M Ribeiro, Georg Zeller, Janet M Thornton

{"title":"Investigating enzyme function by geometric matching of catalytic motifs.","authors":"Raymund E Hackett, Ioannis G Riziotis, Martin Larralde, António J M Ribeiro, Georg Zeller, Janet M Thornton","doi":"10.1002/pro.70744","DOIUrl":"10.1002/pro.70744","url":null,"abstract":"The rapidly growing universe of predicted protein structures offers opportunities for data driven exploration but requires computationally scalable and interpretable tools. We developed a method to detect catalytic features in protein structures, providing insights into enzyme function and mechanism. A library of 6780 3D coordinate sets describing enzyme catalytic sites, referred to as templates, has been collected from manually curated examples of 762 enzyme catalytic mechanisms described in the Mechanism and Catalytic Site Atlas. For template searching we optimized the geometric-matching algorithm Jess. We implemented RMSD and residue orientation filters to differentiate catalytically informative matches from spurious ones. We validated this approach on a non-redundant set of high quality experimental (n = 3751, <40% amino acid identity) enzyme structures with well annotated catalytic sites as well as predicted structures of the human proteome. We show matching catalytic templates solely on structure is more sensitive than sequence- and 3D-structure-based approaches in identifying homology between distantly related enzymes. Since geometric matching does not depend on conserved sequence motifs or even common evolutionary history, we are able to identify examples of structural active site similarity in highly divergent and possibly convergent enzymes. Such examples make interesting case studies into the evolution of enzyme function. Though not intended for characterizing substrate-specific binding pockets, the speed and knowledge-driven interpretability of our method make it well suited for expanding enzyme active-site annotation across large predicted proteomes. We provide the method and template library as a Python module, Enzyme Motif Miner, at https://github.com/rayhackett/enzymm and as a webserver at https://www.ebi.ac.uk/thornton-srv/m-csa/enzymm.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70744"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13448135/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148685520","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Molecular logics in dual sensor regulation of diguanylate cyclase activity-Phosphorylation OR blue-light activation. 二胍酸环化酶活性双传感器调控的分子逻辑——磷酸化或蓝光激活。

IF 5.6 3区生物学

Protein Science Pub Date : 2026-09-01 DOI: 10.1002/pro.70765

Maximilian Fuchs, Andreas Winkler

{"title":"Molecular logics in dual sensor regulation of diguanylate cyclase activity-Phosphorylation OR blue-light activation.","authors":"Maximilian Fuchs, Andreas Winkler","doi":"10.1002/pro.70765","DOIUrl":"10.1002/pro.70765","url":null,"abstract":"Bacterial cells use multiple environmental cues to regulate levels of the second messenger cyclic dimeric GMP. This compound influences key lifestyle decisions such as motility, biofilm formation, or virulence. Although many diguanylate cyclases (DGCs) combined with various sensory domains have been studied previously, how distinct inputs are integrated within individual enzymes remains incompletely understood. Here, we investigate a cyanobacterial family of dual sensor DGCs that combine an N-terminal receiver (Rec) domain followed by a light-oxygen-voltage (LOV) domain upstream of a diguanylate cyclase (GGDEF) domain. Using in vivo activity screening and in vitro characterization, we determined how phosphorylation and blue light, individually and jointly, regulate enzymatic activity. By measuring kinetic parameters across four defined functional states, unphosphorylated or phosphorylated, in combination with dark or light states, we reveal logic gate-like behaviors. One representative, LaRldC, integrates both signals with pronounced fold-changes in activity, consistent with overall OR-type logic and with light acting as the dominant input. Our results demonstrate its function as a molecular gate coupling phosphorylation and illumination sensing to cyclic-di-GMP formation. These findings provide valuable insights into multi-signal decision-making in cyanobacteria and establish further understanding of how modular sensory domains are wired to control bacterial second-messenger signaling.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70765"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13501900/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148809002","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Disulfide engineering of the FANCM-MHF complex reveals constraints on crosslink design in symmetric oligomers. FANCM-MHF配合物的二硫工程揭示了对称低聚物交联设计的约束。

IF 5.6 3区生物学

Protein Science Pub Date : 2026-09-01 DOI: 10.1002/pro.70761

Sho Ito, Tatsuya Nishino

{"title":"Disulfide engineering of the FANCM-MHF complex reveals constraints on crosslink design in symmetric oligomers.","authors":"Sho Ito, Tatsuya Nishino","doi":"10.1002/pro.70761","DOIUrl":"10.1002/pro.70761","url":null,"abstract":"The Fanconi anemia complementation group M protein (FANCM)-MHF complex is required for branched DNA recognition in the Fanconi anemia pathway, but structural analysis of the intact complex has been hindered by dissociation of FANCM from the FANCM-associated histone fold (MHF) heterotetramer under crystallization conditions. Here, we used structure-guided disulfide engineering to stabilize the FANCM-MHF interface and test whether local geometry is sufficient to predict crosslinking specificity in a symmetric oligomeric assembly. Using endogenous FANCM Cys759 as an anchor, we designed two MHF2 variants, Q74C and L77C. Both supported oxidation-dependent crosslinking in the context of the FANCM-MHF complex, but with distinct outcomes. Q74C formed the intended FANCM-MHF2 disulfide, enabled crystallization of the intact heteropentamer, and preserved DNA-binding behavior under the tested conditions. In contrast, L77C favored a competing MHF2-MHF2 disulfide and yielded only the MHF heterotetramer after FANCM dissociation. Structural analysis further showed distinct crosslinking states for the two MHF tetramers in the asymmetric unit, consistent with local conformational heterogeneity at the MHF dimer-dimer interface. These results show that geometric plausibility alone does not predict crosslinking specificity in symmetric oligomers. Instead, symmetry-related competing pathways can redirect the reaction toward an alternative assembly state. This study provides a practical route to stabilizing FANCM-MHF and reveals a key design constraint for engineered disulfides in symmetric multimeric assemblies.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70761"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13479805/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148762610","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A simple probabilistic AlphaFold interaction score. 一个简单的概率AlphaFold交互得分。

IF 5.6 3区生物学

Protein Science Pub Date : 2026-09-01 DOI: 10.1002/pro.70760

Mihaly Badonyi, Agnes Toth-Petroczy

{"title":"A simple probabilistic AlphaFold interaction score.","authors":"Mihaly Badonyi, Agnes Toth-Petroczy","doi":"10.1002/pro.70760","DOIUrl":"10.1002/pro.70760","url":null,"abstract":"AlphaFold has enabled large-scale prediction of protein-protein and protein-nucleic acid complexes, but ranking and assessing the quality of predicted models remain challenging. Existing confidence scores are often highly parametrized and provide limited interpretability. We introduce a simple geometric framework that converts AlphaFold-predicted aligned error (PAE) into conditional contact probability. We show that these probabilities are well calibrated to the fraction of native contacts observed across experimentally determined structures. Motivated by this, we define the Pinc score (Probability of interface native contacts) as the mean contact probability between interacting chains. Because the probabilistic interpretation extends to individual residues, Pinc captures local structural constraint beyond interfacial burial, enabling residue-level prioritization of hotspot positions for mutational studies. Depending solely on a single empirically fixed contact radius, Pinc offers an interpretable path from PAE to interface confidence, matching or exceeding the classification performance of more complex methods across five independent benchmark sets. We provide a portable, dependency-free C program and a Google Colab notebook for calculating Pinc scores for AlphaFold models at https://git.mpi-cbg.de/tothpetroczylab/Pinc.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70760"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13488364/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148797406","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Phase separation leads to significant conformational change in the N-terminal region of the prion protein. 相分离导致朊病毒蛋白n端区域的显著构象变化。

IF 5.6 3区生物学

Protein Science Pub Date : 2026-09-01 DOI: 10.1002/pro.70742

Suman Pal, Jayant B Udgaonkar

{"title":"Phase separation leads to significant conformational change in the N-terminal region of the prion protein.","authors":"Suman Pal, Jayant B Udgaonkar","doi":"10.1002/pro.70742","DOIUrl":"10.1002/pro.70742","url":null,"abstract":"Understanding the link between phase separation (PS) of disease-linked proteins to form liquid-like condensates, and their aggregation, requires insights into the conformational changes that the proteins undergo inside the condensates as they age and become solid-like. In this study, the structural changes undergone by the mouse prion protein (moPrP) inside condensate induced by PS have been characterized. Hydrogen-deuterium exchange in conjunction with mass spectrometry reveals that the N-terminal region (NTR), which is unstructured in monomeric native moPrP, gains significant stable structure as the condensate ages. The structured C-terminal domain remains native-like, albeit with higher stability, but subtle changes are seen. Conformational change initiates in the native monomer inside the condensate, with different regions undergoing rapid, slow, or no conformational change as it ages. The β1-α1 loop undergoes rapid conformational change to lose stability, while the NTR gains structure slowly concomitantly with conformational change at the C-terminal end of α3. Infrared (IR) spectroscopy shows that β-structure forms, IR and circular dichroism spectroscopy indicate that secondary structure becomes heterogeneous, and dynamic light scattering measurements reveal that the protein forms oligomeric nanoscale assemblies as the condensate ages. The formation of the nanoscale assemblies inside the condensate is responsible for the fraction of protein present as mobile monomer decreasing with time of aging, when fluorescence recovery after photobleaching is quantified. Such assembly and the resultant conformational change in the protein appear to be responsible for a change in its material properties of the condensate, which manifests itself as a liquid-like to solid-like transition.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70742"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13440355/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148679762","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Graph identification of proteins in tomograms (GRIP-Tomo) 2.0: Topologically aware classification for proteins. 层析图中蛋白质的图形识别（GRIP-Tomo） 2.0：对蛋白质的拓扑感知分类。

IF 5.6 3区生物学

Protein Science Pub Date : 2026-09-01 DOI: 10.1002/pro.70721

Chengxuan Li, August George, Reece Neff, Doo Nam Kim, Trevor Moser, Kate Baldwin, Malio Nelson, Arsam Firoozfar, James E Evans, Margaret S Cheung

{"title":"Graph identification of proteins in tomograms (GRIP-Tomo) 2.0: Topologically aware classification for proteins.","authors":"Chengxuan Li, August George, Reece Neff, Doo Nam Kim, Trevor Moser, Kate Baldwin, Malio Nelson, Arsam Firoozfar, James E Evans, Margaret S Cheung","doi":"10.1002/pro.70721","DOIUrl":"10.1002/pro.70721","url":null,"abstract":"Cryo-electron tomography (cryo-ET) enables structural characterization of biomolecules under near-native conditions. Existing approaches for interpreting the resulting three-dimensional volumes are computationally expensive and have difficulty interpreting density associated with small proteins/complexes. To explore alternate approaches for identifying proteins in cryo-ET data, we pursued a Graph Network and topologically invariant approach. Here, we report on a fast algorithm that distinguishes volumes containing protein density from noise by searching for nuances of evolutionarily conserved motifs and the geometric characteristics of protein structure. Graph Identification of Proteins in Tomograms (GRIP-Tomo) 2.0 is a machine-learning pipeline that extracts interpretable topological features of protein structures within noisy experimental backgrounds. Compared to version 1.0, the new pipeline includes three upgrades that significantly improve performance, including synthetic tomogram generation simulating realistic noise, graph-based persistent feature extraction as protein fingerprints, and High Performance Computing acceleration. GRIP-Tomo 2.0 achieves over 90% accuracy in distinguishing proteins from noise for synthetic datasets and over 80% accuracy for real datasets with Angstroms per pixel close to 1 from the protein mixtures of in-house samples, which represents a foundational step toward advancing cryo-ET workflows and empowering automated detection of both small and large proteins for visual proteomics. https://github.com/EMSL-Computing/grip-tomo.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 9","pages":"e70721"},"PeriodicalIF":5.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13454385/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148702173","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Correction to "Structure-activity relationship study shows versatility in substrate specificity for heterodisulfide reductase in methanogenic archaea". 更正“构效关系研究显示产甲烷古菌中异二硫还原酶的底物特异性具有多功能性”。

IF 5.6 3区生物学

Protein Science Pub Date : 2026-08-01 DOI: 10.1002/pro.70711

引用次数: 0

Protein Data Bank Japan: A unified portal for integrating structural and chemical data to explore protein-ligand interactions in PDB and PubChem. 日本蛋白质数据库：整合结构和化学数据的统一门户，以探索PDB和PubChem中的蛋白质配体相互作用。

IF 5.6 3区生物学

Protein Science Pub Date : 2026-08-01 DOI: 10.1002/pro.70702

Gert-Jan Bekker, Chioko Nagao, Satomi Niwa, Genji Kurisu

{"title":"Protein Data Bank Japan: A unified portal for integrating structural and chemical data to explore protein-ligand interactions in PDB and PubChem.","authors":"Gert-Jan Bekker, Chioko Nagao, Satomi Niwa, Genji Kurisu","doi":"10.1002/pro.70702","DOIUrl":"10.1002/pro.70702","url":null,"abstract":"Protein Data Bank Japan (https://pdbj.org/) is the Asian hub of three-dimensional (3D) macromolecular structure data and a founding member of the global Protein Data Bank (PDB) network. Over two decades, we have curated and distributed experimentally determined structures, complementing international collaborations with Research Collaboratory for Structural Bioinformatics (RCSB) PDB, Biological Magnetic Resonance Data Bank, Protein Data Bank in Europe (PDBe), and Electron Microscopy Data Bank. In response to user demand for integrated structural and chemical data, we developed a new PubChem Portal that enables interactive exploration of compound-protein interactions. Users can view ligand binding poses in 3D via our Web Graphics Library (WebGL)-based Molmil viewer, with key interactions highlighted and key residues displayed in semi-transparent stick models, enhanced through integration with secondary databases (e.g., Dynamics DB, eF-site) for advanced insights into molecular dynamics and electrostatics. The system supports filtering by UniProt ID, Enzyme Commission (EC) number, Pfam ID, or PROSITE ID to identify structurally related compounds and visualizes protein-ligand interactions. A dynamic two-dimensional (2D) Japan Agency for Medical Research and Development representation enables real-time atom-level navigation, with clickable atoms linking to 3D structures. This tool allows users to explore compound-protein interaction landscapes, identify potential binding modes, and guide experimental design, such as mutagenesis or crystallization. The portal offers a comprehensive, user-centered ecosystem that bridges chemical and structural data, enhancing access to biological insights through integrated visualization and analysis.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 8","pages":"e70702"},"PeriodicalIF":5.6,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13346351/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148412755","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

NEPRO forms fibrous scaffolds in the nucleolus to maintain nucleolar integrity and orchestrate ribosome biogenesis. NEPRO在核仁中形成纤维支架，以维持核仁的完整性和协调核糖体的生物发生。

IF 5.6 3区生物学

Protein Science Pub Date : 2026-08-01 DOI: 10.1002/pro.70710

Abhishek Kumar, Shailesh Kumar Gupta, Yogendra Pratap Mathuria, Buddhi Prakash Jain, Debasish Kumar Ghosh

{"title":"NEPRO forms fibrous scaffolds in the nucleolus to maintain nucleolar integrity and orchestrate ribosome biogenesis.","authors":"Abhishek Kumar, Shailesh Kumar Gupta, Yogendra Pratap Mathuria, Buddhi Prakash Jain, Debasish Kumar Ghosh","doi":"10.1002/pro.70710","DOIUrl":"10.1002/pro.70710","url":null,"abstract":"The nucleolus is a dynamic, membrane-less organelle whose sub-compartmentalization regulates ribosome biogenesis, RNA processing, and stress signaling. While Nucleolus and Neural Progenitor protein (NEPRO) has been classified as a nucleolar protein involved in ribosomopathy, its intra-nucleolar organization, structural features, and functional contributions have remained less understood. Here, we show that NEPRO localizes predominantly to the edges of the nucleolar boundary, forming distinct pools that do not localize to the canonical nucleolar subdomains. NEPRO assembles into elongated fibers in nucleolus. Recombinant NEPRO self-polymerizes into fibrils under mildly acidic conditions, and these fibers also contour the periphery of the dense fibrillar component of nucleolus. NEPRO depletion disrupted nucleolar integrity. NEPRO harbors an arginine-rich nucleolar localization signal (NoLS: 442-460 amino acid region), and interaction of the NoLS of NEPRO with GNL3's acidic C-terminal region nucleates and concentrates NEPRO, lowering its fibrillation threshold and promoting pH-dependent fibrillar assembly in nucleolus. Affinity pulldown of NEPRO's N-terminal coiled-coil domain identified nucleolar interacting proteins enriched in ribosomal and rRNA-processing factors. NEPRO knockdown impaired 40S/60S/80S ribosome assembly, induced G0/G1 arrest, and cell senescence. Collectively, our data define NEPRO as a structural scaffold protein that orchestrates nucleolar structural organization and ribosome biogenesis through formation of fibrous structures, with its dysfunction leading to nucleolar stress and cell-cycle arrest.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"35 8","pages":"e70710"},"PeriodicalIF":5.6,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13347622/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148412793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0