The role of (alpha beta) protomer interaction in determining functional characteristics of red cell Na,K-ATPase.
We have examined the possibility that interaction of (alpha beta) protomers within a diprotomer is responsible for some anomalous characteristics of red cell Na,K-ATPase by examining their response to two inhibitors, FITC and H2DIDS, which bind covalently, and to ouabain, which debinds slowly from red cell pumps. The phenomena we examined were: (1) the biphasic curve relating Na,K-ATPase activity to ATP concentration, and (2) protection of Na pumps against vanadate inhibition by external Na. If interaction of (alpha beta) protomers within a diprotomer were responsible for these phenomena, random inactivation of (alpha beta) protomers should have resulted in a high proportion of (alpha beta) promtomers with an inhibited protomer as a partner, and therefore should have significantly altered the consequences of subunit interaction. With each inhibitor, 60-70% inhibition of ATPase activity did not alter the functional characteristics of the residual activity. We conclude that interaction of functional (alpha beta) protomers does not explain the phenomena which we investigated. This is consistent with our previous observation that Na,K pumps of red cell membranes exist as monomeric (alpha beta) protomers (Martin, D.W. and Sachs, V.R. (1992) J. Biol. Chem. 267, 23922-23929).
Sachs
J R
JR
Department of Medicine, State University of New York at Stony Brook 11794-8151.
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism; D001665:Binding Sites; D004910:Erythrocyte Membrane / Q000201:enzymology*; D016650:Fluorescein-5-isothiocyanate / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D010042:Ouabain / Q000494:pharmacology; D010766:Phosphorylation; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014638:Vanadates / Q000494:pharmacology
D014638:Vanadates; D010042:Ouabain; C018564:dihydro-DIDS; D000255:Adenosine Triphosphate; D000254:Sodium-Potassium-Exchanging ATPase; D016650:Fluorescein-5-isothiocyanate; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid