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.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 在样品处理过程中，尤其是在使用典型的胰酶消化方案时，可能会出现脱酰胺现象。我们已经开发了一种18O标记方法来监控这些人工脱酰胺，其中酶消化是在18O标记的水中进行的。而在体内，由于-NH_2基团被-OH基团取代而导致每个脱酰胺基的+1 Da质量移动，而在消化过程中发生的那些会导致+3 Da的质量移动，额外的+2 Da是由于一个18O原子的加入而产生的。这项研究最近(Li等人，2008)发表在《美国质谱学学会杂志》上，2008，19,855-864。