Goldfinger Lawrence E, Ptak Celeste, Jeffery Erin D, Shabanowitz Jeffrey, Han Jaewon, Haling Jacob R, Sherman Nicholas E, Fox Jay W, Hunt Donald F, Ginsberg Mark H
Division of Rheumatology, Department of Medicine, University of California, San Diego, La Jolla, California 92093, USA.
J Proteome Res. 2007 May;6(5):1806-11. doi: 10.1021/pr060630l. Epub 2007 Apr 17.
We used a TAP-tag approach to identify candidate binding proteins for the related Ras family GTPases: H-Ras, R-Ras, and Rap1A. Protein complexes were isolated from mouse fibroblasts, and component proteins were identified by a combination of nanoflow HPLC and tandem mass spectrometry. H-Ras was found to associate with numerous cytoskeletal proteins including talin-1. R-Ras and Rap1A each associated with various signaling molecules, many of which are membrane-associated. Thus, we have established the first database of potential Ras interactors in mammalian cells.
我们采用TAP标签法来鉴定与相关Ras家族GTP酶(H-Ras、R-Ras和Rap1A)结合的候选蛋白。从小鼠成纤维细胞中分离蛋白质复合物,并通过纳流HPLC和串联质谱联用的方法鉴定其组成蛋白。结果发现H-Ras与包括踝蛋白-1在内的众多细胞骨架蛋白相关联。R-Ras和Rap1A各自与多种信号分子相关联,其中许多信号分子与膜相关。因此,我们建立了第一个哺乳动物细胞中潜在Ras相互作用分子的数据库。