Regulation of chicken gizzard ecto-ATPase activity by modulators that affect its oligomerization status.
The major ectonucleoside triphosphate phosphohydrolase in the chicken gizzard smooth muscle membranes is an ecto-ATPase, an integral membrane glycoprotein belonging to the E-ATPase (or E-NTPDase) family. The gizzard ecto-ATPase is distinguished by its unusual kinetic properties, temperature dependence, and response to a variety of modulators. Compounds that promote oligomerization of the enzyme protein, i.e., concanavalin A, chemical cross-linking agent, and eosin iodoacetamide, increase its activity. Compounds that inhibit some ion-motive ATPases, e.g., sulfhydryl reagents, xanthene derivatives, NBD-halides, and suramin, also inhibit the gizzard ecto-ATPase, but not another E-ATPase, the chicken liver ecto-ATP-diphosphohydrolase, which contains the same conserved regions as the ecto-ATPase. Furthermore, inhibition of the gizzard ecto-ATPase by these compounds as well as detergents is not prevented by preincubation of the membranes with the substrate, ATP, indicating that their interaction with the enzyme occurs at a locus other than the catalytic site. On the other hand, the inhibitory effect of these compounds, except suramin, is abolished or reduced if the membranes are preincubated with concanavalin A. It is concluded that these structurally unrelated modulators exert their effect by interfering with the oligomerization of the ecto-ATPase protein. Our findings suggest that, under physiological conditions, the gizzard smooth muscle ecto-ATPase may exhibit a range of activities determined by membrane events that affect the status of oligomerization of the enzyme.
Caldwell
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CC
Department of Chemistry, San Diego State University, California 92182-1030, USA.
Hornyak
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SC
Pendleton
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Campbell
D
D
Knowles
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AF
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives / Q000494:pharmacology; D002731:4-Chloromercuribenzenesulfonate / Q000494:pharmacology; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D002645:Chickens; D003902:Detergents / Q000494:pharmacology; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic; D005895:Gizzard, Avian / Q000201:enzymology*; D006902:Hydroxymercuribenzoates / Q000494:pharmacology; D008962:Models, Theoretical; D009130:Muscle, Smooth / Q000201:enzymology*; D011487:Protein Conformation; D013498:Suramin / Q000494:pharmacology; D014966:Xanthenes / Q000494:pharmacology
D003902:Detergents; D006902:Hydroxymercuribenzoates; D014966:Xanthenes; C030601:4-hydroxymercuribenzoate; C032930:7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole; D002731:4-Chloromercuribenzenesulfonate; D013498:Suramin; D000251:Adenosine Triphosphatases; C027201:ectoATPase; D009327:4-Chloro-7-nitrobenzofurazan