|ZFIN ID: ZDB-IMAGE-130808-4|
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.
|Acknowledgments:||ZFIN wishes to thank the journal for permission to reproduce figures from this article. Please note that this material may be protected by copyright.|