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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 在样品处理过程中，脱氨酸可能是作为伪影发生的，尤其是在使用典型的胰蛋白酶消化方案时。我们已经开发了一种18o标记的方法来监测这些人工脱氨基，其中酶消化是在18o标记的水中进行的。由于体内脱氨基替代-OH组，导致每个脱氨基的+1 Da质量转移，但在消化过程中发生的-OH组导致+3 da质量转移，而额外的+2 da由于掺入一个18o原子而产生。这项研究最近发表在《美国质谱学会杂志》，2008，19，855-864上。