13-C nuclear magnetic resonance studies on the mechanism of action of carbonic anhydrase.
Binding of the substrate, bicarbonate, to bovine cobalt carbonic anhydrase (carbonate hydrolyase, EC 4.2.1.1) has been studied with 13-C nuclear magnetic resonance. Two binding sites for bicarbonate have been identified. One loosely binds bicarbonate, inhibits p-nitrophenyl acetate activity, and must be the bicarbonate substrate binding site; the other tightly binds bicarbonate, is noninhibitory, and plays another role. Spinlattice relaxation times for the carbon atom of bicarbonate indicate that the substrate bicarbonate is bound directly to the metal center of the enzyme, while the other bicarbonate is bound in the outer coordination sphere of the metal. It is proposed that dehydration proceeds via HCO-3 minus coordinated directly to the metal center, while the outer sphere bicarbonate facilitates catalytically important proton transfers.
D000818:Animals; D001639:Bicarbonates / Q000378:metabolism; D001665:Binding Sites; D002247:Carbon Isotopes; D002256:Carbonic Anhydrases / Q000378:metabolism*; D002417:Cattle; D005583:Fourier Analysis; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy
D001639:Bicarbonates; D002247:Carbon Isotopes; D002256:Carbonic Anhydrases