Characterization of gill (Na+ K)–activated adenosine triphosphatase from chinook salmon, Oncorhynchus tshawytscha

Abstract

(Na ⁺K)-activated ATPase activity from gills of yearling spring chinook was examined using a new rapid assay method. Characterization of the enzyme activity was performed. Optimal activity was obtained at pH 7.2 in the presence of 240 mM NaCl, 120 mM KCl, 20 mM MgCl₂ and 10 mM Na₂ATP. Maximal inhibition of the enzyme was observed in the presence of 0.5 mM ouabain. Differential centrifugation indicated that 75% of the enzymatic activity was sedimented at 1000 × g. Only 8% of the activity was found in the microsomal pellet. Treatment with 0.1% sodium deoxycholate liberated activity from the 1000 × g pellet and elevated the activity of the microsomal pellet from 8% to 51% of the total activity. This treatment caused a loss of 20% of the original activity of the preparation. Statistical analysis of the sampling procedure for gill (Na ⁺K)-activated ATPase activity indicated that there was small variation in the technique itself when compared to variation between the individual gill arches and between individual fish. Results indicate that for meaningful comparisons of groups of fish, the sampling of the gill arches must be standardized and a large number of individual fish must be sampled.