title: The Use of Metal Fluoride Compounds as Phosphate Analogs for Understanding the Structural Mechanism in P-type ATPases.
abstract: The membrane-bound protein family, P-type ATPases, couples ATP hydrolysis with substrate transport across the membrane and forms an obligatory auto-phosphorylated intermediate in the transport cycle. The metal fluoride compounds, BeF x , AlF x , and MgF x , as phosphate analogs stabilize different enzyme structural states in the phosphoryl transfer/hydrolysis reactions, thereby fixing otherwise short-lived intermediate and transient structural states and enabling their biochemical and atomic-level crystallographic studies. The compounds thus make an essential contribution for understanding of the ATP-driven transport mechanism. Here, with a representative member of P-type ATPase, sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA), we describe the method for their binding and for structural and functional characterization of the bound states, and their assignments to states occurring in the transport cycle.
authors.affiliation: Asahikawa Medical University, 2-1-1-1, Midorigaokahigashi, Asahikawa, Hokkaido, 078-8510, Japan.
authors.forename: Stefania J
authors.initials: SJ
authors.lastname: Danko
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: D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism*; D004795:Enzyme Stability; D005459:Fluorides / Q000737:chemistry*; D006868:Hydrolysis; D008670:Metals / Q000737:chemistry; D008967:Molecular Biology / Q000379:methods*; D010710:Phosphates / Q000737:chemistry
chemical_list: D008670:Metals; D010710:Phosphates; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D005459:Fluorides
keywords: Aluminum fluoride (AlF x ); Beryllium fluoride (BeF x ); Ca2+ pump; Magnesium fluoride (MgF x ); Metal fluoride; P-type ATPase; Phosphate analog; Phosphoryl transfer; Phosphorylated intermediate; SERCA (sarco(endo)plasmic reticulum Calcium ATPase)