Fig. S2
- ID
- ZDB-FIG-130808-4
- Publication
- Dagliyan et al., 2013 - Rational design of a ligand-controlled protein conformational switch
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Testing different lengths of double linkers between uniRapR subdomains. (A) Double linker connecting the subdomains-A and -B are shown in red and cyan. The linker with asterisks is the one used in the current version of uniRapR. (B) Src-uniRapR-cerulean-myc constructs were expressed in HEK293T cells. Cell lysates were blotted with anti-GFP to confirm the expression of the construct. In dimerization-based switch (rapR), coexpressed FRB was also tested with anti-GFP. Cell lysates were pulled down with anti-myc and mixed with substrate paxillin in the presence of ATP for 10 min. Reaction suspension was blotted with anti-myc to confirm the binding, with anti–pY31-paxilin to confirm the phosphorylation of the substrate. Linker 2 is the one required for functional uniRapR. (C) The design with truncated linker 2 (linker 1) does not provide control over Src. |