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ISOTOPIC LABELING FOR THE MEASUREMENT OF GLOBAL PROTEIN LEVELS AND TURNOVER IN VIVO

机译:用于测量体内全球蛋白质水平和营业额的同位素标记

摘要

An entire complement or plurality of isotopically labeled amino acids are introduced into the diet of a test subject. Sufficient amounts of the isotopically labeled amino acids are provided to the subject in order to ensure that the subject incorporates a large percentage of isotopically labeled amino acids into newly synthesized proteins. Tissue samples are removed from the subject at different points in time and proteins are extracted and separated so that different proteins of different tissues can be individually analyzed and their amount and pattern of isotopic labeling can be determined. In a preferred embodiment, the methodology can be combined with proteolytic digestion to peptides and analysis by mass spectrometry in order to measure rates of protein turnover in vivo relating to thousands of different proteins.
机译:将完整的补体或多个同位素标记的氨基酸引入测试对象的饮食中。向受试者提供足够量的同位素标记的氨基酸,以确保受试者将大量百分比的同位素标记的氨基酸掺入新合成的蛋白质中。在不同的时间点从受试者身上取出组织样品,提取并分离蛋白质,以便可以分别分析不同组织中的不同蛋白质,并可以确定其同位素标记的数量和方式。在优选的实施方案中,该方法可以与蛋白水解消化成肽并通过质谱分析相结合,以测量体内与数千种不同蛋白质有关的蛋白质周转率。

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