Wellcome Trust Centre for Gene Regulation and Expression, University of Dundee, Dundee, UK.
Nat Methods. 2011 Aug 28;8(10):849-51. doi: 10.1038/nmeth.1679.
We describe an approach for accurate quantitation of global protein dynamics in Caenorhabditis elegans. We adapted stable-isotope labeling with amino acids in cell culture (SILAC) for nematodes by feeding worms a heavy lysine- and heavy arginine-labeled Escherichia coli strain and report a genetic solution to elminate the problem of arginine-to-proline conversion. Combining our approach with quantitative proteomics methods, we characterized the heat-shock response in worms.
我们描述了一种用于精确定量秀丽隐杆线虫中全局蛋白质动力学的方法。我们通过给线虫喂食一种重赖氨酸和重精氨酸标记的大肠杆菌菌株,将稳定同位素标记的氨基酸在细胞培养中的应用(SILAC)方法进行了适应性改造,并报告了一种遗传解决方案来消除精氨酸到脯氨酸转换的问题。我们将这种方法与定量蛋白质组学方法相结合,对蠕虫的热休克反应进行了特征描述。