title: State of aggregation of the (Ca2+ + Mg2+)-ATPase studied using saturation-transfer electron spin resonance.
abstract: The state of aggregation of the (Ca2+ + Mg2+)-ATPase in the membrane of sarcoplasmic reticulum and in reconstituted membrane systems has been studied using saturation-transfer electron spin resonance (ST-ESR). Saturation-transfer ESR spectra show that in the sarcoplasmic reticulum, the ATPase is relatively free to rotate, with an effective rotational correlation time of approx. 33 microseconds at 4 degrees C, consistent with a monomeric or dimeric structure. The rate of rotation is observed to decrease with decreasing molar ratio of lipid to protein. In reconstituted systems, rotational motion of the ATPase on the millisecond time scale ceases when the lipids are in the gel phase. Addition of decavanadate, which causes the formation of crystalline arrays in negatively stained electron micrographs, results in only a small reduction in rotation rate for the ATPase in the membrane. The experiments are interpreted in terms of a short-lived (on the millisecond time scale) protein-protein interaction, with the formation of crystalline clusters of ATPase molecules which form and melt rapidly.
authors.affiliation: Department of Biochemistry, University of Southampton, U.K.
authors.forename: R M
authors.initials: RM
authors.lastname: Napier
authors.forename: J M
authors.initials: JM
authors.lastname: East
authors.forename: A G
authors.initials: AG
authors.lastname: Lee
mesh_terms: D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D003460:Crystallization; D004578:Electron Spin Resonance Spectroscopy; D007425:Intracellular Membranes / Q000201:enzymology; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D009132:Muscles / Q000201:enzymology; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000201:enzymology*; D014638:Vanadates / Q000494:pharmacology
chemical_list: D046911:Macromolecular Substances; D014638:Vanadates; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases