title: Phosphorylation/dephosphorylation of reconstituted shark Na+,K(+)-ATPase: one phosphorylation site per alpha beta protomer.
abstract: In the present investigation reconstitution of Na+,K(+)-ATPase increases the number of phosphorylation sites (EP) of solubilized enzyme from 4.2 +/- 0.3 nmol/mg to 6.9 +/- 0.6 nmol/mg. The latter figure corresponds to one phosphorylation site per alpha beta-promoter. A cholesterol content > 10 mol% in the liposome bilayer and a high extracellular [Na+] are necessary to obtain this high value. Spontaneous dephosphorylation after maximum phosphorylation in Na+ is biphasic both in solubilized enzyme and after reconstitution. The rate of dephosphorylation compares with the specific hydrolytic Na(+)-ATPase activity measured at exactly identical conditions for all three preparations assuming parallel dephosphorylation of at least two phosphointermediates. The distribution of EP-species is found to vary among the three enzyme preparation used, i.e., membrane bound, solubilized, and reconstituted Na+,K(+)-ATPase, however in all the equilibrium is strongly poised away from the E1P-form.
authors.affiliation: Institute of Biophysics, University of Aarhus, Denmark.
authors.forename: F
authors.initials: F
authors.lastname: Cornelius
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002784:Cholesterol / Q000032:analysis; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D046911:Macromolecular Substances; D010766:Phosphorylation; D012754:Sharks; D012964:Sodium / Q000494:pharmacology; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry / Q000378:metabolism*
chemical_list: D008081:Liposomes; D046911:Macromolecular Substances; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D002784:Cholesterol; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase