title: Monomer-dimer association constant of solubilized sarcoplasmic reticulum ATPase.
abstract: The monomer-dimer association constant of solubilized and delipidated sarcoplasmic reticulum ATPase was measured by large zone elution gel chromatography in the presence of a high concentration (18.6 mM) of the nonionic detergent dodecyloctaethylene glycol monoether (C12E8) and of different ATPase protein concentrations in the range of 0.74 (6.4 nM monomers) to 30 (0.26 microM monomers) microgram/ml. The association equilibrium constant (Ka) obtained from the concentration-dependent dissociation curve was 9.37 X 10(7) M-1 at 24 degrees C. The derived free energy change (delta G0) for the monomer-dimer association was -10.8 kcal/mo, reflecting a high degree of tightness between inter-subunit domains in soluble dimeric ATPase. A steep dissociation curve within a short natural logarithmic span (2.5 units) was obtained when the degree of dissociation increased from 0.1 to 0.9, suggesting that a conformational drift accompanies the dissociation of soluble dimeric ATPase. A unique leading boundary was formed in the large zone chromatographies, indicating a reversible equilibrium which was rapid when compared to the time taken for the chromatographic run. Enzymatic activity was continuously monitored in the eluate, revealing that soluble ATPase at different degrees of dissociation was active.
authors.forename: J L
authors.initials: JL
authors.lastname: Silva
authors.forename: S
authors.initials: S
authors.lastname: Verjovski-Almeida
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D001386:Azides / Q000378:metabolism; D002850:Chromatography, Gel / Q000379:methods; D046911:Macromolecular Substances; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*; D012995:Solubility
chemical_list: C026397:3'(O)-(3-(N-(4-azido-2-nitrophenyl)amino)propionyl)-8-azidoadenosine 5'-triphosphate; D001386:Azides; D046911:Macromolecular Substances; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases