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 资助的中心拨款提供的资源的众多研究子项目之一。对子项目和子项目主要研究者的主要支持可能是由其他来源提供的，包括其他 NIH 来源。 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金额，而不是 NCRR 拨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，isoAsp 残基也可以通过 Asp 异构化形成，尽管其发生速度要慢得多。 Asp 和 isoAsp 残基的分化很重要，因为后者通常会导致蛋白质构象和功能发生更显着的变化，这与许多蛋白质错误折叠疾病和其他病理过程有关。本实验室建立了基于电子捕获解离 (ECD)-MS/MS 的方法，其中来自 isoAsp 残基的诊断性 c+57/z+-57 离子用于两种异构体形式的区分和相对定量。该方法已应用于研究以下几个模型系统中的 Asn 脱酰胺和 Asp 异构化。 (1) 开发自上而下的同天冬氨酸组学研究方法： (2) 在自下而上的 isoAsp 分析中使用内切蛋白酶 Glu-C： (3) N端isoAsp鉴定： (4)利用ECD和ETD区分谷氨酰胺脱酰胺产物： 该研究发表于《分析化学》，2010, 82, 3606-3615。 (5) ECD 和 ETD 对 β 连接肽的异常断裂： 该研究发表在《美国质谱学会杂志》，2011, 22, 480-491 上。