In-vitro/In-vivo Translation Platform Group , GlaxoSmithKline , 1250 S Collegeville Road , Collegeville , Pennsylvania 19426 , United States.
Department of Chemistry and Biochemistry , Brigham Young University , Provo , Utah 84604 , United States.
Anal Chem. 2019 Aug 6;91(15):9732-9740. doi: 10.1021/acs.analchem.9b01329. Epub 2019 Jul 15.
We describe an analytical strategy allowing for the direct quantification of stable isotope label incorporation in newly synthesized proteins following administration of the stable isotope tracer deuterium oxide. We present a demonstration of coupling high-resolution mass spectrometry, metabolic stable isotope labeling, and MS/MS-based isotopologue quantification for the measurement of protein turnover. Stable isotope labeling with deuterium oxide, followed by immonium ion isotopologue quantification, is a more sensitive strategy for determining protein fractional synthesis rates compared to peptide centric mass isotopomer distribution analysis approaches when labeling time and/or stable isotope tracer exposure is limited and, as such, offers a great advantage for human studies.
我们描述了一种分析策略，允许在给予氘水稳定同位素示踪剂后直接定量新合成蛋白质中的稳定同位素标记掺入。我们展示了一种将高分辨率质谱、代谢稳定同位素标记和基于 MS/MS 的同量异位素定量相结合的方法，用于测量蛋白质周转率。与基于肽的质量同位素分布分析方法相比，氘水稳定同位素标记后进行铵离子同量异位素定量是一种更灵敏的策略，用于确定蛋白质的合成率，特别是在标记时间和/或稳定同位素示踪剂暴露有限的情况下，因此，对于人体研究具有很大的优势。
