USE OF 18O LABELS TO MONITOR DEAMIDATION DURING SAMPLE PROCESSING

使用 18O 标签监测样品处理过程中的脱酰胺作用

• 批准号: 7955974
• 负责人: PETER B. O'CONNOR
• 金额: $ 1.18万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7955974
• 关键词: American; Biology; Computer Retrieval of Information on Scientific Projects Database; Digestion; Funding; Grant; Institution; Journals; Label; Lead; Mass Spectrum Analysis; Medicine; Monitor; Morphologic artifacts; Process; Protocols documentation; Publishing; Research; Research Personnel; Resources; Sampling; Site; Societies; Source; United States National Institutes of Health; Water; deamidation; in vivo

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Deamidation may occur as an artifact during sample processing, particularly when using typical tryptic digestion protocols. We have developed an 18O labeling approach to monitor these artificial deamidations, where the enzymatic digestion was carried out in 18O labeled water. While in vivo deamidation causes a +1 Da mass shift per deamidation site due to the substitution of the -NH2 group by the -OH group, those occurred during digestion lead to a +3 Da mass shift, with the extra +2 Da arising from the incorporation of one 18O atom. This study has recently been published (Li et al., 2008) in the Journal of the American Society for Mass Spectrometry, 2008, 19, 855-864.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 在样品处理过程中，脱酰胺可能会作为伪影发生，特别是在使用典型的胰蛋白酶消化方案时。我们开发了一种 18O 标记方法来监测这些人工脱酰胺，其中酶消化是在 18O 标记水中进行的。虽然体内脱酰胺作用由于 -NH2 基团被 -OH 基团取代而导致每个脱酰胺位点产生 +1 Da 质量位移，但消化过程中发生的那些导致 +3 Da 质量位移，其中由于一个 18O 原子的掺入而产生额外的 +2 Da 质量位移。这项研究最近发表在美国质谱学会杂志 (Journal of the American Society for Mass Spectrometry), 2008, 19, 855-864 上 (Li et al., 2008)。