title: Protein kinase C phosphorylates plasma membrane Ca2+ pump isoform 4a at its calmodulin binding domain.
abstract: Phosphorylation by protein kinase C of isoform 4a of the human plasma membrane Ca2+ pump (hPMCA4a) was studied using the COS cell expression system. Phosphorylation of several truncated mutants of hPMCA4a indicated that a single phosphorylation site lies in a region between residues 1113 and 1125. This region is within the calmodulin binding domain and contains a single phosphorylatable residue, serine 1115. Converting this serine to an alanine diminished phosphorylation greatly. Phosphorylation, done in the absence of calmodulin, did not affect subsequent calmodulin binding, but previous binding of calmodulin did inhibit phosphorylation. Moreover, no significant shift in the calmodulin response curve of hPMCA4a was observed when phosphorylation was mimicked by converting serine 1115 to an acidic residue. The calmodulin binding domain of hPMCA4a is much longer than other calmodulin binding domains and has been suggested to consist of two binding lobes interrupted by a short nonbinding region. The findings of this study indicate that serine 1115 is the residue phosphorylated by protein kinase C, and that it lies within the nonbinding region of the calmodulin binding domain.
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Foundation, Rochester, Minnesota 55905, USA.
authors.forename: A K
authors.initials: AK
authors.lastname: Verma
authors.forename: K
authors.initials: K
authors.lastname: Paszty
authors.forename: A G
authors.initials: AG
authors.lastname: Filoteo
authors.forename: J T
authors.initials: JT
authors.lastname: Penniston
authors.forename: A
authors.initials: A
authors.lastname: Enyedi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017510:Protein Folding; D011493:Protein Kinase C / Q000378:metabolism*
chemical_list: D002147:Calmodulin; D007527:Isoenzymes; D011493:Protein Kinase C; D000252:Calcium-Transporting ATPases