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在稳定同位素标记的氨基酸细胞培养 (SILAC) 条件下进行细胞培养时,以细胞系非依赖性的方式防止精氨酸到脯氨酸的转化。

Preventing arginine-to-proline conversion in a cell-line-independent manner during cell cultivation under stable isotope labeling by amino acids in cell culture (SILAC) conditions.

机构信息

Functional Proteome Analysis, German Cancer Research Center, 69120 Heidelberg, Germany.

出版信息

Anal Biochem. 2011 May 1;412(1):123-5. doi: 10.1016/j.ab.2011.01.011. Epub 2011 Jan 15.

Abstract

Quantitative proteomics has increasingly gained impact in life science research as a tool to describe changes in protein expression between different cellular states. Stable isotope labeling by amino acids in cell culture (SILAC) is a powerful technique for relative quantification of proteins. However, the accuracy of quantification is impaired by the metabolic conversion of arginine to proline resulting in additional heavy labeled proline peptide satellites. Here we reinvestigated the addition of unlabeled proline during cell cultivation under SILAC conditions considering several thousand peptides and demonstrated that the arginine-to-proline conversion is prevented independent of the cell line used.

摘要

定量蛋白质组学作为一种描述不同细胞状态下蛋白质表达变化的工具,在生命科学研究中越来越受到重视。稳定同位素标记的氨基酸在细胞培养中的应用(SILAC)是一种相对定量蛋白质的强大技术。然而,由于精氨酸代谢转化为脯氨酸导致额外的重标记脯氨酸肽卫星,从而影响了定量的准确性。在这里,我们重新研究了在 SILAC 条件下细胞培养过程中添加未标记脯氨酸的情况,考虑了几千个肽,并证明了无论使用何种细胞系,都可以防止精氨酸到脯氨酸的转化。

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