Figure 9.

Membrane binding, Triton X-100 sensitivity, and coimmunoprecipitation of fatty acylated and farnesylated Fyn constructs. (A) COS-1 cells were transfected with cDNAs encoding wild-type Fyn, FynKRas, or G₂A,C₃S-FynKRas. Transfected cells were radiolabeled with Tran³⁵S-label, homogenized, and fractionated as described in Materials and Methods. Distribution into the P100 fraction was quantitated by phosphorimaging. Values represent the mean of two experiments. (B) COS-1 cells were transfected as described in A, followed by fractionation into 1% Triton X-100 soluble (S) or resistant (R) fractions. Distribution of Fyn constructs into the TX100 resistant fraction was quantitated after immunoprecipitation by Western blotting analysis, as described in Materials and Methods. Values represent the mean of two experiments. (C) COS-1 cells were transfected as described in A and coimmunoprecipitation analysis of Fyn protein with CD8-ζ was performed as described in Fig. 7 C, legend.