title: Phosphorylation/Dephosphorylation Assays.
abstract: The P-type ATPases form an autophosphorylated intermediate with ATP, and its isomeric transition and hydrolysis are obligatory events in the ATP-driven pump and thus for the energy coupling. The analyses of these reactions are therefore crucial for understanding the mechanism of the pump function and diseases caused by its defects. Here we describe the methods to analyze these processes in the transport cycle with a representative member of P-type ATPase family, SERCA1a, sarco(endo)plasmic reticulum Ca(2+)-ATPase.
authors.affiliation: Asahikawa Medical University, 2-1-1-1, Midorigaokahigashi, Asahikawa, Hokkaido, 078-8510, Japan. hisuzuki@asahikawa-med.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Suzuki
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D008967:Molecular Biology / Q000379:methods*; D010766:Phosphorylation; D011817:Rabbits; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000255:Adenosine Triphosphate; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D002118:Calcium
keywords: ATP; ATP -driven pumps; Aspartyl phosphate; Autoradiography; Inorganic phosphate; P-type ATPase s; Phosphorylated intermediate s; Phosphorylation; SDS -PAGE; SERCA