Site-Specific Hydrogen Deuterium Exchange Difference Mass Spectrometry Measurements for Ligand Binding

IF 2.7 2区 化学 Q2 BIOCHEMICAL RESEARCH METHODS
Joseph Anacleto, , , Suzanne Ackloo, , , Cheryl Arrowsmith, , , Esther Wolf, , , Yves Leblanc, , , Cristina Lento, , and , Derek J. Wilson*, 
{"title":"Site-Specific Hydrogen Deuterium Exchange Difference Mass Spectrometry Measurements for Ligand Binding","authors":"Joseph Anacleto,&nbsp;, ,&nbsp;Suzanne Ackloo,&nbsp;, ,&nbsp;Cheryl Arrowsmith,&nbsp;, ,&nbsp;Esther Wolf,&nbsp;, ,&nbsp;Yves Leblanc,&nbsp;, ,&nbsp;Cristina Lento,&nbsp;, and ,&nbsp;Derek J. Wilson*,&nbsp;","doi":"10.1021/jasms.5c00229","DOIUrl":null,"url":null,"abstract":"<p >Conventional bottom-up HDX-MS experiments are highly suitable for use in drug development; however, a major limitation of this approach is that it generally provides only peptide-level structural resolution. Site specific (<i>i.e</i>., single amino acid-resolved) HDX-MS measurements have been achieved using ECD/ETD, but the low efficiency of these fragmentation techniques, combined with poor ion transmission associated with ‘detuning’ the instrument to fully prevent deuterium scrambling, results in sensitivity losses that make ligand binding measurements impractical in a ‘real-world’ (<i>e.g</i>., drug development) context. Here we apply a recently developed method for zero scrambling, high efficiency ECD in the challenging context of ligand binding differential HDX experiments, demonstrating a wealth of additional information that can be acquired when HDX-MS analyses are conducted at the amino acid level.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"36 10","pages":"2315–2318"},"PeriodicalIF":2.7000,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/jasms.5c00229","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of the American Society for Mass Spectrometry","FirstCategoryId":"92","ListUrlMain":"https://pubs.acs.org/doi/10.1021/jasms.5c00229","RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0

Abstract

Conventional bottom-up HDX-MS experiments are highly suitable for use in drug development; however, a major limitation of this approach is that it generally provides only peptide-level structural resolution. Site specific (i.e., single amino acid-resolved) HDX-MS measurements have been achieved using ECD/ETD, but the low efficiency of these fragmentation techniques, combined with poor ion transmission associated with ‘detuning’ the instrument to fully prevent deuterium scrambling, results in sensitivity losses that make ligand binding measurements impractical in a ‘real-world’ (e.g., drug development) context. Here we apply a recently developed method for zero scrambling, high efficiency ECD in the challenging context of ligand binding differential HDX experiments, demonstrating a wealth of additional information that can be acquired when HDX-MS analyses are conducted at the amino acid level.

配体结合的位点特异性氢氘交换差质谱测量。
传统的自下而上的HDX-MS实验非常适合用于药物开发;然而,这种方法的一个主要限制是,它通常只提供肽水平的结构分辨率。使用ECD/ETD已经实现了位点特异性(即单氨基酸分解)的HDX-MS测量,但是这些碎片化技术的低效率,加上与“失谐”仪器相关的离子传输不良,无法完全防止氘的混乱,导致灵敏度损失,使得配体结合测量在“现实世界”(例如药物开发)环境中不切实际。在这里,我们在配体结合差异HDX实验的挑战性背景下应用了一种最近开发的零干扰、高效ECD方法,展示了在氨基酸水平上进行HDX- ms分析时可以获得的丰富的附加信息。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
CiteScore
5.50
自引率
9.40%
发文量
257
审稿时长
1 months
期刊介绍: The Journal of the American Society for Mass Spectrometry presents research papers covering all aspects of mass spectrometry, incorporating coverage of fields of scientific inquiry in which mass spectrometry can play a role. Comprehensive in scope, the journal publishes papers on both fundamentals and applications of mass spectrometry. Fundamental subjects include instrumentation principles, design, and demonstration, structures and chemical properties of gas-phase ions, studies of thermodynamic properties, ion spectroscopy, chemical kinetics, mechanisms of ionization, theories of ion fragmentation, cluster ions, and potential energy surfaces. In addition to full papers, the journal offers Communications, Application Notes, and Accounts and Perspectives
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信