DIFFERENTIATION OF ISOMERIC AMINO ACID RESIDUES IN PEPTIDES USING ECD

使用 ECD 区分肽中的异构氨基酸残基

• 批准号: 8365528
• 负责人: CHI-WEI LIN
• 金额: $ 5.08万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-08-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365528
• 关键词: American; Amino Acids; Analytical Chemistry; Asparagine; Biological Models; Biology; Development; Diagnostic; Disease; Dissociation; Electrons; Funding; Glutamine; Grant; Ions; Isoaspartic Acid; Journals; Laboratories; Link; Mass Spectrum Analysis; Medicine; Methods; N-terminal; National Center for Research Resources; Pathologic Processes; Peptides; Physiological; Principal Investigator; Protein Conformation; Publishing; Relative (related person); Research; Research Infrastructure; Resources; Societies; Source; Succinimides; United States National Institutes of Health; base; cost; deamidation; protein function; protein misfolding

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Under physiological conditions, asparagine (Asn) residues can deamidate spontaneously, generating a mixture of aspartic (Asp) and isoaspartic acid (isoAsp) residues via a succinimide intermediate. In addition, isoAsp residue may also be formed through Asp isomerization, although this occurs at a much slower rate. Differentiation of Asp and isoAsp residues is important as the latter often causes more significant changes in protein conformation and functions, which has been linked to many protein misfolding diseases and other pathological processes. An electron capture dissociation (ECD)-MS/MS based method was established in this laboratory, where diagnostic c+57/z+-57 ions from isoAsp residues were used for the differentiation and relative quantification of the two isomeric forms. This method has been applied to study the Asn deamidation and Asp isomerization in several model systems below. (1) Development of a top-down approach for isoaspartomics research: (2) Use of the endoprotease Glu-C in bottom-up isoAsp analysis: (3) N-terminal isoAsp identification: (4) Differentiation of glutamine deamidation products using ECD and ETD: This research has been published in Analytical Chemistry, 2010, 82, 3606-3615. (5) Unusual fragmentation of beta-linked peptides by ECD and ETD: This research has been published in the Journal of the American Society for Mass Spectrometry, 2011, 22, 480-491.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和子项目主要研究者的主要支持可能由其他来源提供，包括其他NIH来源。 为子项目列出的总成本可能代表子项目使用的中心基础设施的估计数量，而不是NCRR赠款向子项目或子项目工作人员提供的直接资金。 在生理条件下，天冬酰胺（Asn）残基可以自发脱酰胺，通过琥珀酰亚胺中间体生成天冬氨酸（Asp）和异天冬氨酸（isoAsp）残基的混合物。此外，也可以通过Asp异构化形成isoAsp残基，尽管这以慢得多的速率发生。Asp和isoAsp残基的区分是重要的，因为后者经常引起蛋白质构象和功能的更显著变化，这与许多蛋白质错误折叠疾病和其他病理过程有关。本实验室建立了一种基于电子捕获解离（ECD）-MS/MS的方法，利用isoAsp残基的诊断性c+57/z+-57离子对两种异构体进行区分和相对定量。该方法已被应用于以下几个模型系统中的Asn脱酰胺和Asp异构化的研究。 (1)开发自上而下的方法进行同工酶组学研究： (2)内切蛋白酶Glu-C在自下而上isoAsp分析中的用途： (3)N-末端isoAsp鉴别： (4)使用ECD和ETD区分谷氨酰胺脱酰胺产物： 该研究已发表在Analytical Chemistry，2010，82，3606-3615中。 (5)通过ECD和ETD进行的β-连接肽的异常断裂： 该研究已发表在Journal of the American Society for Mass Spectrometry，2011，22，480-491中。