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Figure 6 | Cell Communication and Signaling

Figure 6

From: Complex formation of EphB1/Nck/Caskin1 leads to tyrosine phosphorylation and structural changes of the Caskin1 SH3 domain

Figure 6

Tyrosine phosphorylation leads to structural changes in the Caskin1 SH3 domain. (a) The SH3 domain of Caskin1 was expressed, purified, and phosphorylated in vitro by EphB1 as described in the Methods. The upper panel demonstrates the phosphorylation of the SH3 domain probed with anti-phosphotyrosine antibody. The lower panel shows the protein samples stained with Coomassie blue. (b) Far-UV CD spectra were measured of the unphosphorylated and the phosphorylated SH3 domains. The CD spectrum of the buffer itself is also indicated. (c) Near-UV CD spectra were measured of the unphosphorylated and the phosphorylated SH3 domains. The CD spectrum of the buffer itself is also indicated. These results are representative of three experiments. (d) The SH3 domain of Caskin1 was expressed, purified, and phosphorylated in vitro by EphB1 as described in the Methods. The upper panel demonstrates the time course of phosphorylation of the SH3 domain stained with Coomassie-blue. After blotting the proteins to nitrocellulose membrane, immunoblots were performed using anti-His-tag and anti-phosphotyrosine antibodies (middle and lower panels).

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