Characterisation of Ca2+ or Mg(2+)-dependent nucleoside triphosphatase from rat mesenteric small arteries.
When isolated rat mesenteric small arteries were submitted to 2 s of sonication, a nucleoside triphosphatase activity was released to the medium, mainly from the plasma membrane of the vascular smooth muscle cells. The activity was kinetically characterized: It hydrolysed ATP, UTP and GTP with the same substrate affinity and the same specific activity. CaATP, as well as MgATP were substrates for the enzyme with an apparent Km in the micromolar range. ATPase inhibitors: ouabain, vanadate, AlF4-, oligomycin and N-ethylmaleimide were without effect on the hydrolytic activity. Among other modifiers tested only N,N'-dicyclohexylcarbodiimide caused significant (greater than 30%) inhibition. In the presence of micromolecular concentrations of Ca2+ and Mg2+, small (less than 20 mM) concentrations of Na+, K+, Rb+, Cs+ and choline+, irrespective of the nature of the anion, activated the hydrolysis with an equilibrium ordered pattern, but concentrations of monovalent cation salts above 20 mM decreased the hydrolysis rate. No activation by monovalent cation salts was seen at millimolar concentrations of divalent cations and substrate. On the basis of the results a standard mixture is proposed, which allows a sensitive assay of the specific enzyme activity.
Plesner
L
L
Institute of Biophysics, University of Aarhus, Denmark.
Juul
B
B
Skriver
E
E
Aalkjaer
C
C
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002412:Cations; D002413:Cations, Divalent; D006160:Guanosine Triphosphate / Q000378:metabolism; D006868:Hydrolysis; D008274:Magnesium / Q000378:metabolism*; D008297:Male; D008638:Mesenteric Arteries / Q000201:enzymology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D043583:Nucleoside-Triphosphatase; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry*; D051381:Rats; D011919:Rats, Inbred Strains; D013379:Substrate Specificity; D014544:Uridine Triphosphate / Q000378:metabolism
D002412:Cations; D002413:Cations, Divalent; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D010744:Phosphoric Monoester Hydrolases; D000251:Adenosine Triphosphatases; D043583:Nucleoside-Triphosphatase; D008274:Magnesium; D002118:Calcium; D014544:Uridine Triphosphate