[Energy characteristics of an ATP-hydrolase reaction catalyzed by solubilized Ca2+,Mg2+-ATPase from smooth muscle cell membrane].
The effects of temperature, dielectric permeability and ionic strength on the activity of purified Ca2+, Mg(2+)-ATPase solubilized from myometrial sarcolemma have been studied under saturation of the enzyme with Ca2+, Mg2+ and ATP. The values of activation energy calculated from Arrhenius plots for both ATP hydrolase reactions catalysed by solubilized and reconstituted into azolectin liposomes Ca2+, Mg(2+)-ATPase and Mg2+, ATP-dependent Ca2+ transport by the reconstituted enzyme were 56.4 +/- 1.5, 68.0 +/- 5.1 and 63.1 +/- 2.9 kJ/mol, respectively. Analysis of experimental data in terms of the Laidler-Scatchard and Bronsted-Bjerrum theories revealed that the separation of the reaction products--the chelate MgADP complex--from the active site of the enzyme bearing one unity positive charge is the limiting step of the Ca2+, Mg(2+)-dependent enzymatic ATP-hydrolysis under conditions of substrate saturation. The values of the electrostatic components of the free energy, enthalpy and entropy of activation of the ATP hydrolase reaction were 46.6 +/- 0.3 kJ/mol, -(20.5 +/- 0.4) kJ/mol and -(214.2 +/- 4.3) J/(mol.degrees K), respectively. The nonelectrostatic component of activation enthalpy was 76.9 kJ/mol. The results obtained suggest that changes in polarity of the incubation medium markedly affect the activity of transport Ca2+, Mg(2+)-ATPase solubilized from smooth muscle cell plasma membranes and that the electrostatic interactions between the enzyme active site and specific reagents (MgADP, in particular) significantly contribute to the energetics of the ATP hydrolase reaction.
Kosterin
S A
SA
Slinchenko
N N
NN
Gergalova
G L
GL
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D001692:Biological Transport; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002384:Catalysis; D002462:Cell Membrane / Q000201:enzymology; D004560:Electricity; D004734:Energy Metabolism; D004789:Enzyme Activation; D005260:Female; D006867:Hydrolases / Q000378:metabolism*; D006868:Hydrolysis; D009215:Myometrium / Q000201:enzymology*; D009994:Osmolar Concentration; D012995:Solubility; D013552:Swine; D013696:Temperature; D013816:Thermodynamics
D000255:Adenosine Triphosphate; D006867:Hydrolases; D017301:Ca(2+) Mg(2+)-ATPase; D002118:Calcium
Energeticheskie kharakteristiki ATP-gidrolaznoĭ reaktsii, kataliziruemoĭ soliubilizirovannoĭ Ca2+,Mg2+-ATPazoĭ plazmaticheskoĭ membrany gladkomyshechnykh kletok.