title: A single site (Ser16) phosphorylation in phospholamban is sufficient in mediating its maximal cardiac responses to beta -agonists.
abstract: Phospholamban (PLB) can be phosphorylated at Ser(16) by cyclic AMP-dependent protein kinase and at Thr(17) by Ca(2+)-calmodulin-dependent protein kinase during beta-agonist stimulation. A previous study indicated that mutation of S16A in PLB resulted in lack of Thr(17) phosphorylation and attenuation of the beta-agonist stimulatory effects in perfused mouse hearts. To further delineate the functional interplay between dual-site PLB phosphorylation, we generated transgenic mice expressing the T17A mutant PLB in the cardiac compartment of the null background. Lines expressing similar levels of T17A mutant, S16A mutant, or wild-type PLB in the null background were characterized in parallel. Cardiac myocyte basal mechanics and Ca(2+) kinetics were similar among the three groups. Isoproterenol stimulation was associated with phosphorylation of both Ser(16) and Thr(17) in wild-type PLB and Ser(16) phosphorylation in T17A mutant PLB, whereas there was no detectable phosphorylation of S16A mutant PLB. Phosphorylation of Ser(16) alone in T17A mutant PLB resulted in responses of the mechanical and Ca(2+) kinetic parameters to isoproterenol similar to those in wild-type myocytes, which exhibited dual-site PLB phosphorylation. However, those parameters were significantly attenuated in the S16A mutant myocytes. Thus, Ser(16) in PLB can be phosphorylated independently of Thr(17) in vivo, and phosphorylation of Ser(16) is sufficient for mediating the maximal cardiac responses to beta-adrenergic stimulation.
authors.affiliation: Department of Pharmacology & Cell Biophysics, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267-0575, USA.
authors.forename: G
authors.initials: G
authors.lastname: Chu
authors.forename: J W
authors.initials: JW
authors.lastname: Lester
authors.forename: K B
authors.initials: KB
authors.lastname: Young
authors.forename: W
authors.initials: W
authors.lastname: Luo
authors.forename: J
authors.initials: J
authors.lastname: Zhai
authors.forename: E G
authors.initials: EG
authors.lastname: Kranias
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D006321:Heart / Q000187:drug effects / Q000502:physiology*; D007545:Isoproterenol / Q000494:pharmacology*; D007700:Kinetics; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D009206:Myocardium / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000235:genetics; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D011943:Receptors, Adrenergic, beta / Q000502:physiology*; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D012694:Serine; D000099211:Phospholamban
chemical_list: D002118:Calcium; D002135:Calcium-Binding Proteins; D000252:Calcium-Transporting ATPases; D007545:Isoproterenol; D018995:Myosin Heavy Chains; D011943:Receptors, Adrenergic, beta; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D012694:Serine; D000099211:Phospholamban