Protein Science最新文献_第3页

Mesoscale explorer: Visual exploration of large‐scale molecular models 中尺度探索者大尺度分子模型的可视化探索

IF 8 3区生物学

Protein Science Pub Date : 2024-09-18 DOI: 10.1002/pro.5177

Alexander Rose, David Sehnal, David S. Goodsell, Ludovic Autin

{"title":"Mesoscale explorer: Visual exploration of large‐scale molecular models","authors":"Alexander Rose, David Sehnal, David S. Goodsell, Ludovic Autin","doi":"10.1002/pro.5177","DOIUrl":"https://doi.org/10.1002/pro.5177","url":null,"abstract":"The advent of cryo‐electron microscopy (cryo‐EM) and cryo‐electron tomography (cryo‐ET), coupled with computational modeling, has enabled the creation of integrative 3D models of viruses, bacteria, and cellular organelles. These models, composed of thousands of macromolecules and billions of atoms, have historically posed significant challenges for manipulation and visualization without specialized molecular graphics tools and hardware. With the recent advancements in GPU rendering power and web browser capabilities, it is now feasible to render interactively large molecular scenes directly on the web. In this work, we introduce Mesoscale Explorer, a web application built using the Mol* framework, dedicated to the visualization of large‐scale molecular models ranging from viruses to cell organelles. Mesoscale Explorer provides unprecedented access and insight into the molecular fabric of life, enhancing perception, streamlining exploration, and simplifying visualization of diverse data types, showcasing the intricate details of these models with unparalleled clarity.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"51 1","pages":""},"PeriodicalIF":8.0,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253063","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

O‐GlcNAc modification of HSP27 alters its protein interactions and promotes refolding of proteins through the BAG3/HSP70 co‐chaperone HSP27 的 O-GlcNAc 修饰会改变其与蛋白质的相互作用，并通过 BAG3/HSP70 协同伴侣促进蛋白质的再折叠

IF 8 3区生物学

Protein Science Pub Date : 2024-09-18 DOI: 10.1002/pro.5173

Afraah Javed, Oleta T. Johnson, Aaron T. Balana, Regan F. Volk, Andreas Langen, Benjamin S. Ahn, Balyn W. Zaro, Jason E. Gestwicki, Matthew R. Pratt

{"title":"O‐GlcNAc modification of HSP27 alters its protein interactions and promotes refolding of proteins through the BAG3/HSP70 co‐chaperone","authors":"Afraah Javed, Oleta T. Johnson, Aaron T. Balana, Regan F. Volk, Andreas Langen, Benjamin S. Ahn, Balyn W. Zaro, Jason E. Gestwicki, Matthew R. Pratt","doi":"10.1002/pro.5173","DOIUrl":"https://doi.org/10.1002/pro.5173","url":null,"abstract":"Almost all types of cellular stress induce post‐translational O‐GlcNAc modifications of proteins, and this increase promotes cell survival. We previously demonstrated that O‐GlcNAc on certain small heat shock proteins (sHSPs), including HSP27, directly increases their chaperone activity as one potential protective mechanism. Here, we furthered our use of synthetic proteins to prepare biotinylated sHSPs and show that O‐GlcNAc modification of HSP27 also changes how it interacts within the sHSP system and the broader HSP network. Specifically, we show that O‐GlcNAc modified HSP27 binds more strongly to the co‐chaperone protein BAG3, which then promotes refolding of a model substrate by HSP70. We use proteomics to identify other potential HSP27 interactions that are changed by O‐GlcNAc, including one that we confirm with another sHSP, αB‐crystallin. These findings add additional evidence for O‐GlcNAc as a switch for regulating protein–protein interactions and for modifications of chaperones as one mechanism by which O‐GlcNAc protects against protein aggregation.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"54 1","pages":"e5173"},"PeriodicalIF":8.0,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253061","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

MeCP2 is a naturally supercharged protein with cell membrane transduction capabilities MeCP2 是一种具有细胞膜传导能力的天然超荷蛋白

IF 8 3区生物学

Protein Science Pub Date : 2024-09-14 DOI: 10.1002/pro.5170

Alexander V. Beribisky, Anna Huber, Victoria Sarne, Andreas Spittler, Nyamdelger Sukhbaatar, Teresa Seipel, Franco Laccone, Hannes Steinkellner

{"title":"MeCP2 is a naturally supercharged protein with cell membrane transduction capabilities","authors":"Alexander V. Beribisky, Anna Huber, Victoria Sarne, Andreas Spittler, Nyamdelger Sukhbaatar, Teresa Seipel, Franco Laccone, Hannes Steinkellner","doi":"10.1002/pro.5170","DOIUrl":"https://doi.org/10.1002/pro.5170","url":null,"abstract":"The intrinsically disordered protein MeCP2 is a global transcriptional regulator encoded by the MECP2 gene. Although the structured domains of MeCP2 have been the subject of multiple studies, its unstructured regions have not been that extensively characterized. In this work, we show that MeCP2 possesses properties akin to those of supercharged proteins. By utilizing its unstructured portions, MeCP2 can successfully transduce across cell membranes and localize to heterochromatic foci in the nuclei, displaying uptake levels a third lower than a MeCP2 construct fused to the cell‐penetrating peptide TAT. MeCP2 uptake can further be enhanced by the addition of compounds that promote endosomal escape following cellular trafficking by means of macropinocytosis. Using a combination of in silico prediction algorithms and live‐cell imaging experiments, we mapped the sequence in MeCP2 responsible for its cellular incorporation, which bears a striking resemblance to TAT itself. Transduced MeCP2 was shown to interact with HDAC3. These findings provide valuable insight into the properties of MeCP2 and may be beneficial for devising future protein‐based treatment strategies.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"31 1","pages":""},"PeriodicalIF":8.0,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253087","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

HER4 is a high‐affinity dimerization partner for all EGFR/HER/ErbB family proteins HER4 是所有表皮生长因子受体/HER/ErbB 家族蛋白的高亲和性二聚化伙伴

IF 8 3区生物学

Protein Science Pub Date : 2024-09-14 DOI: 10.1002/pro.5171

Pradeep Kumar Singh, Soyeon Kim, Adam W. Smith

{"title":"HER4 is a high‐affinity dimerization partner for all EGFR/HER/ErbB family proteins","authors":"Pradeep Kumar Singh, Soyeon Kim, Adam W. Smith","doi":"10.1002/pro.5171","DOIUrl":"https://doi.org/10.1002/pro.5171","url":null,"abstract":"Human epidermal growth factor receptors (HER)—also known as EGFR or ErbB receptors—are a subfamily of receptor tyrosine kinases (RTKs) that play crucial roles in cell growth, division, and differentiation. HER4 (ErbB4) is the least studied member of this family, partly because its expression is lower in later stages of development. Recent work has suggested that HER4 can play a role in metastasis by regulating cell migration and invasiveness; however, unlike EGFR and HER2, the precise role that HER4 plays in tumorigenesis is still unresolved. Early work on HER family proteins suggested that there are direct interactions between the four members, but to date, there has been no single study of all four receptors in the same cell line with the same biophysical method. Here, we quantitatively measure the degree of association between HER4 and the other HER family proteins in live cells with a time‐resolved fluorescence technique called pulsed interleaved excitation fluorescence cross‐correlation spectroscopy (PIE‐FCCS). PIE‐FCCS is sensitive to the oligomerization state of membrane proteins in live cells, while simultaneously measuring single‐cell protein expression levels and diffusion coefficients. Our PIE‐FCCS results demonstrate that HER4 interacts directly with all HER family members in the cell plasma membrane. The interaction between HER4 and other HER family members intensified in the presence of a HER4‐specific ligand. Our work suggests that HER4 is a preferred dimerization partner for all HER family proteins, even in the absence of ligands.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"211 1","pages":""},"PeriodicalIF":8.0,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253088","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

BracketMaker: Visualization and optimization of chemical protein synthesis BracketMaker：化学蛋白质合成的可视化和优化

IF 8 3区生物学

Protein Science Pub Date : 2024-09-14 DOI: 10.1002/pro.5174

Judah L. Evangelista, Michael S. Kay

{"title":"BracketMaker: Visualization and optimization of chemical protein synthesis","authors":"Judah L. Evangelista, Michael S. Kay","doi":"10.1002/pro.5174","DOIUrl":"https://doi.org/10.1002/pro.5174","url":null,"abstract":"Chemical protein synthesis (CPS), in which custom peptide segments of ~20–60 aa are produced by solid‐phase peptide synthesis and then stitched together through sequential ligation reactions, is an increasingly popular technique. The workflow of CPS is often depicted with a “bracket” style diagram detailing the starting segments and the order of all ligation, desulfurization, and/or deprotection steps to obtain the product protein. Brackets are invaluable tools for comparing multiple possible synthetic approaches and serve as blueprints throughout a synthesis. Drawing CPS brackets by hand or in standard graphics software, however, is a painstaking and error‐prone process. Furthermore, the CPS field lacks a standard bracket format, making side‐by‐side comparisons difficult. To address these problems, we developed BracketMaker, an open‐source Python program with built‐in graphic user interface (GUI) for the rapid creation and analysis of CPS brackets. BracketMaker contains a custom graphics engine which converts a text string (a protein sequence annotated with reaction steps, introduced herein as a standardized format for brackets) into a high‐quality vector or PNG image. To aid with new syntheses, BracketMaker's “AutoBracket” tool automatically performs retrosynthetic analysis on a set of segments to draft and rank all possible ligation orders using standard native chemical ligation, protection, and desulfurization techniques. AutoBracket, in conjunction with an improved version of our previously reported Automated Ligator (Aligator) program, provides a pipeline to rapidly develop synthesis plans for a given protein sequence. We demonstrate the application of both programs to develop a blueprint for 65 proteins of the minimal Escherichia coli ribosome.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"19 1","pages":""},"PeriodicalIF":8.0,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253068","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Molecular mechanics studies of factors affecting overall rate in cascade reactions: Multi‐enzyme colocalization and environment 影响级联反应总速率因素的分子力学研究：多酶共定位和环境

IF 8 3区生物学

Protein Science Pub Date : 2024-09-14 DOI: 10.1002/pro.5175

Shivansh Kaushik, Ta I Hung, Chia‐en A. Chang

{"title":"Molecular mechanics studies of factors affecting overall rate in cascade reactions: Multi‐enzyme colocalization and environment","authors":"Shivansh Kaushik, Ta I Hung, Chia‐en A. Chang","doi":"10.1002/pro.5175","DOIUrl":"https://doi.org/10.1002/pro.5175","url":null,"abstract":"Millions of years of evolution have optimized many biosynthetic pathways by use of multi‐step catalysis. In addition, multi‐step metabolic pathways are commonly found in and on membrane‐bound organelles in eukaryotic biochemistry. The fundamental mechanisms that facilitate these reaction processes provide strategies to bioengineer metabolic pathways in synthetic chemistry. Using Brownian dynamics simulations, here we modeled intermediate substrate transportation of colocalized yeast–ester biosynthesis enzymes on the membrane. The substrate acetate ion traveled from the pocket of aldehyde dehydrogenase to its target enzyme acetyl‐CoA synthetase, then the substrate acetyl CoA diffused from Acs1 to the active site of the next enzyme, alcohol‐O‐acetyltransferase. Arranging two enzymes with the smallest inter‐enzyme distance of 60 Å had the fastest average substrate association time as compared with anchoring enzymes with larger inter‐enzyme distances. When the off‐target side reactions were turned on, most substrates were lost, which suggests that native localization is necessary for efficient final product synthesis. We also evaluated the effects of intermolecular interactions, local substrate concentrations, and membrane environment to bring mechanistic insights into the colocalization pathways. The computation work demonstrates that creating spatially organized multi‐enzymes on membranes can be an effective strategy to increase final product synthesis in bioengineering systems.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"2 1","pages":""},"PeriodicalIF":8.0,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253069","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Analysis of EGFR binding hotspots for design of new EGFR inhibitory biologics 分析表皮生长因子受体结合热点，设计新型表皮生长因子受体抑制生物制剂

IF 8 3区生物学

Protein Science Pub Date : 2024-09-14 DOI: 10.1002/pro.5141

Claiborne W. Tydings, Bhuminder Singh, Adam W. Smith, Kaitlyn V. Ledwitch, Benjamin P. Brown, Christine M. Lovly, Allison S. Walker, Jens Meiler

{"title":"Analysis of EGFR binding hotspots for design of new EGFR inhibitory biologics","authors":"Claiborne W. Tydings, Bhuminder Singh, Adam W. Smith, Kaitlyn V. Ledwitch, Benjamin P. Brown, Christine M. Lovly, Allison S. Walker, Jens Meiler","doi":"10.1002/pro.5141","DOIUrl":"https://doi.org/10.1002/pro.5141","url":null,"abstract":"The epidermal growth factor (EGF) receptor (EGFR) is activated by the binding of one of seven EGF‐like ligands to its ectodomain. Ligand binding results in EGFR dimerization and stabilization of the active receptor conformation subsequently leading to activation of downstream signaling. Aberrant activation of EGFR contributes to cancer progression through EGFR overexpression/amplification, modulation of its positive and negative regulators, and/or activating mutations within EGFR. EGFR targeted therapeutic antibodies prevent dimerization and interaction with endogenous ligands by binding the ectodomain of EGFR. However, these antibodies have had limited success in the clinic, partially due to EGFR ectodomain resistance mutations, and are only applicable to a subset of patients with EGFR‐driven cancers. These limitations suggest that alternative EGFR targeted biologics need to be explored for EGFR‐driven cancer therapy. To this end, we analyze the EGFR interfaces of known inhibitory biologics with determined structures in the context of endogenous ligands, using the Rosetta macromolecular modeling software to highlight the most important interactions on a per‐residue basis. We use this analysis to identify the structural determinants of EGFR targeted biologics. We suggest that commonly observed binding motifs serve as the basis for rational design of new EGFR targeted biologics, such as peptides, antibodies, and nanobodies.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"211 1","pages":""},"PeriodicalIF":8.0,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253065","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Ligand‐induced CaMKIIα hub Trp403 flip, hub domain stacking, and modulation of kinase activity 配体诱导的 CaMKIIα 中枢 Trp403 翻转、枢纽结构域堆叠和激酶活性调节

IF 8 3区生物学

Protein Science Pub Date : 2024-09-14 DOI: 10.1002/pro.5152

Dilip Narayanan, Anne Sofie G. Larsen, Stine Juul Gauger, Ruth Adafia, Rikke Bartschick Hammershøi, Louise Hamborg, Jesper Bruus‐Jensen, Nane Griem‐Krey, Christine L. Gee, Bente Frølund, Margaret M. Stratton, John Kuriyan, Jette Sandholm Kastrup, Annette E. Langkilde, Petrine Wellendorph, Sara M. Ø. Solbak

{"title":"Ligand‐induced CaMKIIα hub Trp403 flip, hub domain stacking, and modulation of kinase activity","authors":"Dilip Narayanan, Anne Sofie G. Larsen, Stine Juul Gauger, Ruth Adafia, Rikke Bartschick Hammershøi, Louise Hamborg, Jesper Bruus‐Jensen, Nane Griem‐Krey, Christine L. Gee, Bente Frølund, Margaret M. Stratton, John Kuriyan, Jette Sandholm Kastrup, Annette E. Langkilde, Petrine Wellendorph, Sara M. Ø. Solbak","doi":"10.1002/pro.5152","DOIUrl":"https://doi.org/10.1002/pro.5152","url":null,"abstract":"γ‐Hydroxybutyric acid (GHB) analogs are small molecules that bind competitively to a specific cavity in the oligomeric CaMKIIα hub domain. Binding affects conformation and stability of the hub domain, which may explain the neuroprotective action of some of these compounds. Here, we describe molecular details of interaction of the larger‐type GHB analog 2‐(6‐(4‐chlorophenyl)imidazo[1,2‐b]pyridazine‐2‐yl)acetic acid (PIPA). Like smaller‐type analogs, PIPA binding to the CaMKIIα hub domain promoted thermal stability. PIPA additionally modulated CaMKIIα activity under sub‐maximal CaM concentrations and ultimately led to reduced substrate phosphorylation. A high‐resolution X‐ray crystal structure of a stabilized CaMKIIα (6x mutant) hub construct revealed details of the binding mode of PIPA, which involved outward placement of tryptophan 403 (Trp403), a central residue in a flexible loop close to the upper hub cavity. Small‐angle X‐ray scattering (SAXS) solution structures and mass photometry of the CaMKIIα wild‐type hub domain in the presence of PIPA revealed a high degree of ordered self‐association (stacks of CaMKIIα hub domains). This stacking neither occurred with the smaller compound 3‐hydroxycyclopent‐1‐enecarboxylic acid (HOCPCA), nor when Trp403 was replaced with leucine (W403L). Additionally, CaMKIIα W403L hub was stabilized to a larger extent by PIPA compared to CaMKIIα hub wild type, indicating that loop flexibility is important for holoenzyme stability. Thus, we propose that ligand‐induced outward placement of Trp403 by PIPA, which promotes an unforeseen mechanism of hub domain stacking, may be involved in the observed reduction in CaMKIIα kinase activity. Altogether, this sheds new light on allosteric regulation of CaMKIIα activity via the hub domain.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"39 1","pages":""},"PeriodicalIF":8.0,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253090","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Integration of proteomic data with genome‐scale metabolic models: A methodological overview 蛋白质组数据与基因组尺度代谢模型的整合：方法概述

IF 8 3区生物学

Protein Science Pub Date : 2024-09-14 DOI: 10.1002/pro.5150

Farid Zare, Ronan M. T. Fleming

{"title":"Integration of proteomic data with genome‐scale metabolic models: A methodological overview","authors":"Farid Zare, Ronan M. T. Fleming","doi":"10.1002/pro.5150","DOIUrl":"https://doi.org/10.1002/pro.5150","url":null,"abstract":"The integration of proteomics data with constraint‐based reconstruction and analysis (COBRA) models plays a pivotal role in understanding the relationship between genotype and phenotype and bridges the gap between genome‐level phenomena and functional adaptations. Integrating a generic genome‐scale model with information on proteins enables generation of a context‐specific metabolic model which improves the accuracy of model prediction. This review explores methodologies for incorporating proteomics data into genome‐scale models. Available methods are grouped into four distinct categories based on their approach to integrate proteomics data and their depth of modeling. Within each category section various methods are introduced in chronological order of publication demonstrating the progress of this field. Furthermore, challenges and potential solutions to further progress are outlined, including the limited availability of appropriate in vitro data, experimental enzyme turnover rates, and the trade‐off between model accuracy, computational tractability, and data scarcity. In conclusion, methods employing simpler approaches demand fewer kinetic and omics data, consequently leading to a less complex mathematical problem and reduced computational expenses. On the other hand, approaches that delve deeper into cellular mechanisms and aim to create detailed mathematical models necessitate more extensive kinetic and omics data, resulting in a more complex and computationally demanding problem. However, in some cases, this increased cost can be justified by the potential for more precise predictions.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"22 1","pages":""},"PeriodicalIF":8.0,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253064","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Engineering and physicochemical characterization of a novel, stable, symmetric bispecific antibody with dual target‐binding using a common light chain 一种新型、稳定、对称的双特异性抗体的工程设计和理化表征--利用共同的轻链实现双目标结合

IF 8 3区生物学

Protein Science Pub Date : 2024-09-14 DOI: 10.1002/pro.5121

Seiji Saito, Makoto Nakayama, Kaori Yamazaki, Yuya Miyamoto, Keiko Hiraishi, Daisuke Tomioka, Sayaka Takagi‐Maeda, Katsuaki Usami, Nobuaki Takahashi, Shinji Nara, Eiichiro Imai

{"title":"Engineering and physicochemical characterization of a novel, stable, symmetric bispecific antibody with dual target‐binding using a common light chain","authors":"Seiji Saito, Makoto Nakayama, Kaori Yamazaki, Yuya Miyamoto, Keiko Hiraishi, Daisuke Tomioka, Sayaka Takagi‐Maeda, Katsuaki Usami, Nobuaki Takahashi, Shinji Nara, Eiichiro Imai","doi":"10.1002/pro.5121","DOIUrl":"https://doi.org/10.1002/pro.5121","url":null,"abstract":"Bispecific antibodies (BsAbs) have emerged as a major class of antibody therapeutics owing to their substantial potential in disease treatment. While several BsAbs have been successfully approved in recent years, ongoing development efforts continue to focus on optimizing various BsAbs tailored to particular antigens and action mechanisms, aiming to achieve favorable physicochemical properties. BsAbs generally encounter challenges due to their unfavorable physicochemical characteristics and poor manufacturing efficiencies, highlighting the need for optimization to achieve reliable productivity and developability. Herein, we describe the development of a novel symmetric BsAb, REGULGENT™ (N‐term/C‐term), comprising two Fab domains, using a common light chain. The heavy chain fragment encoded two antigen‐binding determinants in one chain. The design and production of REGULGENT™ (N‐term/C‐term) are simple owing to the use of the same light chain, which does not induce heavy and light chain mispairing, frequently observed with the asymmetric BsAb format. REGULGENT™ (N‐term/C‐term) exhibited high expression and low aggregation characteristics during cell culture and stress treatment under low pH conditions. Differential scanning calorimetric data indicated that REGULGENT™ molecules had high conformational stability, similar to that of stabilized monoclonal antibodies. Surface plasmon resonance data showed that REGULGENT™ (N‐term/C‐term) could bind to two antigens simultaneously and exhibited a high affinity for two antigens. In summary, the symmetric BsAb format of REGULGENT™ confers its desirable IgG‐like physicochemical properties, thus making it an excellent candidate for commercial development. The findings demonstrate a novel BsAb with substantial development potential for clinical applications.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"14 1","pages":""},"PeriodicalIF":8.0,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253071","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0