Abstract:
:Membrane preparations from two independent ouabain-resistant HeLa cell clones, HI-B1 and HI-C1, each appear to contain two species of (Na,K)ATPase. Two-thirds of the total (Na,K)ATPase in each mutant is indistinguishable from the enzyme in preparations of wild type cells with respect to ouabain binding, ouabain inhibition of (Na,K)ATPase activity, and dependence of ATP hydrolysis on Na, Mg, K, and ATP concentration. The remaining (Na,K)ATPase activity in the mutants is up to 1000 and 10 000 times, respectively, more resistant to ouabain than wild type enzyme. Resistance results from a lower affinity of the mutant enzymes for the inhibitor. The presence of Na, K, or Mg has little or no effect on the degree of resistance expressed by the mutant enzymes, although the resistance of the wild type enzyme varies 400-fold in the presence of different ligands. Incubation with 5 X 10(-8) M ouabain abolishes the activity of the wild type enzyme without affecting the activity of the resistant enzymes. Using this procedure we compared the parameters of ATP hydrolysis via the resistant and wild type enzymes. Ouabain-resistant (Na,K)ATPase of HI-C1 has an apparent K0.5 for potassium 3-4 times higher than that of either wild type enzyme or the resistant enzyme of HI-B1.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Robbins AR,Baker RMdoi
10.1021/bi00642a600subject
Has Abstractpub_date
1977-11-15 00:00:00pages
5163-8issue
23eissn
0006-2960issn
1520-4995journal_volume
16pub_type
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