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Purification and properties of trout gill AMP deaminase

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Summary

The AMP deaminase has been purified 450–500 fold from 20,000 g supernatants from trout gill. The procedure comprised cellulose phosphate and DEAE-cellulose chromatography. The gill appeared to contain different isoenzymes as indicated by different chromatographic behaviour on cellulose phosphate and different heat stabilities. The two major isoenzymes were compared with respect to their pH optima and the effect of temperature, ATP and inorganic phosphate. The pH optimum is about pH 6.7 at low substrate concentration. A second optimum is found in phosphate buffer. The substrate saturation curve is hyperbolic, even in the absence of KCl or ATP. ATP is an activator of the enzyme in the absence of KCl, but is without effect in the presence of monovalent cations. Among the monovalent cations tested, Na+ is the most potent activator followed by K+ and NH +4 . Inorganic phosphate is an inhibitor of gill AMP deaminase increasing the affinity for its substrate but having no effect on the maximal velocity or the Hill coefficient. The inhibition by phosphate is partially reversed by ATP. ADP and GTP are competitive inhibitors of the enzyme. In addition, the enzyme showed negative cooperativity in the presence of ATP or GTP.

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  1. Laboratoire de Physiologie comparée des Régulations, CNRS, 23, rue du Loess, B.P. 20 CR, F-67037, Strasbourg-Cédex, France

    Jean-P. Raffin

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Raffin, JP. Purification and properties of trout gill AMP deaminase. J Comp Physiol B 154, 55–63 (1984). https://doi.org/10.1007/BF00683216

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