气候变化领域集成服务门户(CCPortal): Structural basis for differential recognition of phosphohistidine-containing peptides by 1-pHis and 3-pHis monoclonal antibodies

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DOI	10.1073/pnas.2010644118
	Structural basis for differential recognition of phosphohistidine-containing peptides by 1-pHis and 3-pHis monoclonal antibodies
	Kalagiri R.; Stanfield R.L.; Meisenhelder J.; la Clair J.J.; Fuhs S.R.; Wilson I.A.; Hunter T.
发表日期	2021
ISSN	00278424
卷号	118 期号:6
英文摘要	In 2015, monoclonal antibodies (mAbs) that selectively recognize the 1-pHis or 3-pHis isoforms of phosphohistidine were developed by immunizing rabbits with degenerate Ala/Gly peptides containing the nonhydrolyzable phosphohistidine (pHis) analog- phosphotriazolylalanine (pTza). Here, we report structures of five rabbit mAbs bound to cognate pTza peptides: SC1-1 and SC50-3 that recognize 1-pHis, and their 3-pHis-specific counterparts, SC39-4, SC44-8, and SC56-2. These cocrystal structures provide insights into the binding modes of the pTza phosphate group that are distinct for the 1- and 3-pHis mAbs with the selectivity arising from specific contacts with the phosphate group and triazolyl ring. The mode of phosphate recognition in the 3-pHis mAbs recapitulates the Walker A motif, as present in kinases. The complementarity-determining regions (CDRs) of four of the Fabs interact with the peptide backbone rather than peptide side chains, thus conferring sequence independence, whereas SC44-8 shows a proclivity for binding a GpHAGA motif mediated by a sterically complementary CDRL3 loop. Specific hydrogen bonding with the triazolyl ring precludes recognition of pTyr and other phosphoamino acids by these mAbs. Kinetic binding experiments reveal that the affinity of pHis mAbs for pHis and pTza peptides is submicromolar. Bound pHis mAbs also shield the pHis peptides from rapid dephosphorylation. The epitope-paratope interactions illustrate how these anti-pHis antibodies are useful for a wide range of research techniques and this structural information can be utilized to improve the specificity and affinity of these antibodies toward a variety of pHis substrates to understand the role of histidine phosphorylation in healthy and diseased states. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Histidine phosphorylation; Phosphohistidine antibodies; Posttranslational modifications
语种	英语
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/180706
作者单位	Molecular and Cell Biology Laboratory, Salk Institute for Biological Studies, San Diego, CA 92037, United States; Department of Integrative Structural and Computational Biology, Scripps Research Institute, San Diego, CA 92037, United States; Department of Chemistry and Biochemistry, University of California San Diego, San Diego, CA 92093, United States; Jack H. Skirball Center for Chemical Biology and Proteomics, Salk Institute for Biological Studies, San Diego, CA 92037, United States; Skaggs Institute for Chemical Biology, Scripps Research Institute, San Diego, CA 92037, United States; Genomics Institute of the Novartis Research Foundation, San Diego, CA 92121, United States
推荐引用方式 GB/T 7714	Kalagiri R.,Stanfield R.L.,Meisenhelder J.,et al. Structural basis for differential recognition of phosphohistidine-containing peptides by 1-pHis and 3-pHis monoclonal antibodies[J],2021,118(6).
APA	Kalagiri R..,Stanfield R.L..,Meisenhelder J..,la Clair J.J..,Fuhs S.R..,...&Hunter T..(2021).Structural basis for differential recognition of phosphohistidine-containing peptides by 1-pHis and 3-pHis monoclonal antibodies.Proceedings of the National Academy of Sciences of the United States of America,118(6).
MLA	Kalagiri R.,et al."Structural basis for differential recognition of phosphohistidine-containing peptides by 1-pHis and 3-pHis monoclonal antibodies".Proceedings of the National Academy of Sciences of the United States of America 118.6(2021).