title: The influence of Mg2+ on anion binding to sarcoplasmic reticulum membranes as detected by 35Cl-NMR.
abstract: 35Cl-NMR spectroscopy has been used to study the competition between anions, including nucleotides, on skeletal muscle sarcoplasmic reticulum membranes. Different chloride binding sites can be distinguished according to their Mg2+ sensitivity. Phosphate binding is enhanced by Mg2+ whereas the anion transport inhibitor pyridoxalphosphate-6-azophenyl-2'-sulfonic acid (PPAPS) binding is not. The affinity of the enzyme for the Mg-adenylyl imidodiphosphate (MgAMP-PNP) complex is decreased whereas that for MgATP is increased. Three sets of binding sites can be discriminated from which chloride is displaced by different anions with varying efficiency. High affinity binding of AMP-PNP and PPAPS occurs at the same site, that can also be occupied by phosphate. Low-affinity binding of PPAPS and AMP-PNP also coincides, but in a site where phosphate binding is negligible. ATP and ADP bind to both sites. In the presence of Mg2+ a third anion binding site can be occupied by phosphate but neither by AMP-PNP nor PPAPS.
authors.affiliation: Institut für Biophysikalische Chemie und Biochemie, Johann-Wolfgang-Goethe-Universität, Frankfurt am Main Germany.
authors.forename: S T
authors.initials: ST
authors.lastname: Janetzky
authors.forename: H
authors.initials: H
authors.lastname: Hanssum
authors.forename: G
authors.initials: G
authors.lastname: Spatz-Kümbel
authors.forename: H G
authors.initials: HG
authors.lastname: Bäumert
mesh_terms: D000249:Adenosine Monophosphate / Q000378:metabolism; D000818:Animals; D000838:Anions / Q000378:metabolism*; D001665:Binding Sites; D066298:In Vitro Techniques; D008274:Magnesium / Q000502:physiology*; D009682:Magnetic Resonance Spectroscopy; D009711:Nucleotides / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D000838:Anions; D009711:Nucleotides; D010710:Phosphates; C057654:pyridoxal phosphate-6-azophenyl-2'-sulfonic acid; D000249:Adenosine Monophosphate; D011732:Pyridoxal Phosphate; D008274:Magnesium