Figure 4. Tyrosines 246 and 293 are required to hold GIT1 in a closed conformation.
(A,C,D,F) COS7 cells were cotransfected with the indicated constructs, lysed, and immunoprecipitated with anti-GFP (A,C,F) or anti-FLAG antibodies (D). Filters with immunoprecipitates (IP) and lysates were blotted to reveal the full length GFP-tagged constructs (with anti-GFP), HA-GIT1-N (with mouse 12CA5 mAb), or GIT1-C2-LZ fragment (with anti-FLAG). (B,E,G) Schemes of the interactions proposed from the experiments presented in (A), (C,D), and (F), respectively. The asterisks in (E) indicate the monomeric mutated carboxy-terminal fragment that can not dimerize. See the Results for details.