Products/Services Used | Details | Operation |
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PCR Cloning and Subcloning> | Codon-optimized cDNAs encoding Nterminal HA or Flag-tagged WT PEAK3 and HA-tagged PEAK3 mutants were synthesized by Genscript and cloned into the Eco RI restriction sites of the pMIG–green fluorescent protein (GFP) Express vector, also known as pMSCVIRES-GFP (34). Short hairpin RNAs (shRNAs) or single-guide RNAs(sgRNAs) were synthesized by Genscript and cloned into pLKO-Tet-On or lentiGuide-Puro (Addgene, catalog no.52963) vectors. | Get A Quote |
Gene Synthesis> | Get A Quote |
The pseudokinase scaffolds PEAK1 and PEAK2 are implicated in cancer cell migration and metastasis. We characterized the regulation and role of the third family member PEAK3 in cell signaling. Similar to PEAK1 and PEAK2, PEAK3 formed both homotypic and heterotypic complexes. In addition, like PEAK1, it bound to the adaptors Grb2 and CrkII. However, unlike PEAK1 and PEAK2, homodimerized PEAK3 also interacted with the ARF GTPase-activating protein ASAP1, the E3 ubiquitin ligase Cbl, and the kinase PYK2. Dimerization and subsequent phosphorylation on Tyr24, likely by a Src family kinase, were required for the binding of PEAK3 to Grb2 and ASAP1. Interactions with Grb2, CrkII, ASAP1, Cbl, and PYK2 exhibited contrasti... More