Reciprocal Regulation of the Platelet-Derived Growth Factor Receptor-β and G Protein-Coupled Receptor Kinase 5 by Cross-Phosphorylation: Effects on Catalysis
Mol Pharmacol Cai et al. 75: 626 Data Supplement
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- Data Supplement - Supplementary Fig. I. PDGFRΒ target phosphorylation sites within GRK5. A, Three GRK5 mutant constructs are depicted diagrammatically, with the locations of Tyr-to-Phe mutations designated by asterisks within the general 3-domain architecture of GRK5. B, HEK 293 cells were co-transfected with plasmids encoding the PDGFRΒ and the indicated GRK5 construct or no protein (empty vector, “None”). As described for Fig. 2, cells were serum-starved and then exposed to serum-free medium lacking or containing 2 nM PDGF-BB, as indicated, and then subjected to GRK5 IP; divided samples were subjected to parallel SDS-PAGE and IB for either GRK5 or phospho-Tyr (pTyr). Tyrosyl phosphorylation of GRK5 was quantitated as in Fig. 2, and plotted as the means ± S.E. of 4 independent experiments. All cell lines expressed equivalent levels of PDGFRΒ, as assessed by flow cytometry (see Methods; data not shown). Compared with the 4YF GRK5: *, p
