Figure 2
From: Dual-faced SH3BGRL: oncogenic in mice, tumor suppressive in humans
mSH3BGRL enhances bind to inactive c-Src and promote c-Src activation. (a) SH3BGRL immunoprecipitates from the various stable cell lines were immunoblotted with antibodies against SH3BGRL, c-Src, p-c-Src Y527 and p-c-Src Y416. (b) Immunoprecipitation of p-c-Src Y527 and p-c-Src Y416 by overexpression of chicken c-Src mutants K295M and Y527F and detected with the indicated antibodies as in (a). (c) Western blots of p-c-Src Y527 and p-c-Src Y416 in cells overexpressing mSH3BGRL with the indicated antibodies in (a). (d) Cells stably expressing vector (Vec), SH3BGRL-P64A (P64A) or SH3BGRL-WT (SH3BGRL) were immunoblotted with the indicated antibodies. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) served as a loading control, and the averaged relative protein expression level is quantified and shown under the particular panels. (e) Immunoprecipitation of p-c-Src Y527 and p-c-Src Y416 by overexpression of wild-type mSH3BGRL and its P64A mutants in CHO cells and detected with the indicated antibodies. (f) Schematic model for SH3BGRL-mediated c-Src activation. The SH3-binding domain from SH3BGRL competes for and displaces the inhibitory SH3-binding SH2 kinase linker region in c-Src, triggering conformational changes that promote c-Src kinase activity.
