Syp Y279, Y304 Can Mediate the Binding of Bcr-Abl to Grb2 and Other Proteins.
In this study we obtained 3 mutants of Syp cDNA Y279F, Y304F and Y546F by using the method of site-directed mutation in vitro. We then inserted them into the pXM mammalian expression vector, and transferred the plasmids into K562 cells. Western Blot has found WT/ 3MT Syp are expressed in the K562 cells. Immunoprecipitation and Immunoblot have found that WT, Y279F, Y304F and Y546F Syp can bind directly to the Bcr-Abl in vivo. However, we found that mutation of Syp Y279F can block the binding of Grb2 SH2 to Syp mutation of Syp Y304F can block the binding of Shc SH2 to Syp in vitro. As a adaptor, Syp can mediate binding of Bcr-Abl to Shc and Grb2.Grb2 bind to Syp pY279, Shc bind to Syp pY304 in vitro.
Zhu
Kui
K
Clinic Lab./ Research Center, Changhai Hospital, Second Military Medical University, Shanghai 200433, China. zhuk@cesmtp.ccf.org
Liu
Xiao-Ping
XP
Zeng
Grant
G
Ye
Fang-Yun
FY