Identification of PDZ Domain Containing Proteins Interacting with 1.2 and PMCA4b
Figure 1
Expression of PDZ array ligands Cav1.2 and PMCA4b. (C-terminal ligands for class I PDZ domains). (a) Determination of C-terminal consensus sequence mammalian Cav1.2 and PMCA4b (b) The final 10 amino acid residues of Cav1.2 and the last 15 amino acid residues of PMCA4b were expressed with a 6xHistidine tag linked by a poly-Glycine linker through insertion into the expression vector pEXP. Cav1.2 () and PMCA4b (), * = stop, possess different C-terminal ends and resulting in interactions with different PDZ domain containing proteins. (c) Tricine-SDS-PAGE to verify the expression of recombinant protein ligands in BL21 bacteria. The shifts between the lanes pEXP (~9 kDa), pEXP-Cav1.2 (8.47 kDa), and pEXP-PMCA4b (8.97 kDa), where the read-through product, pEXP, is larger than the PDZ ligands, confirmed their successful expression. Each lane contained 4 μL of bacterial lysate.
