ENZYMIC CHARACTERISTICS OF ECTO-ADENOSINE TRIPHOSPHATASE IN RAT EPIDIDYMAL INTACT SPERMATOZOA

Ecto-ATPase in rat cauda-epididymal intact spermatozoa has a high degree of substrate specificity for the hydrolysis of ATP and dATP rather than of ADP, AMP, GTP, dGTP, CTP, dCTP, TTP and UTP. The enzyme is activated by bivalent metal ions in the order Mg2+ > Mn2+ > Co2+ > Ca2+. The apparent Km values of the enzyme for Mg2+, Mn2+, Co2+ and Ca2+ are approximately 80, 100, 100 and 150 .mu.M, respectively. Addition of Ca2+ (0.1 or 1 mM) gives no further stimulation of the Mg2+-activated ecto-ATPase activity. The apparent Km value of the enzyme for ATP is 95 .mu.M. Pi (16 mM) inhibits the enzymic activity (by 25%), whereas Na+ (50 mM) or K+ (10 mM) alone or in combination, polyamines (spermine and spermidine; 1-12.5 mM) and nucleic acids (yeast RNA and calf thymus DNA; 0.12 or 0.62 mg/ml) had no significant effect on the activity of the enzyme. Orthovanadate at a relatively low concentration (20 .mu.M) strongly inhibits (.apprx. 50%) the ecto-ATPase activity. Vanadate inhibition can be reversed by noradrenaline [norepinephrine] (2.5 mM). The vanadate-sensitivity of the enzyme increases markedly during spermatozoal maturation in the epididymis. The activity of the spermatozoal ecto-ATPase decreases progressively during the epididymal transit of the testicular spermatozoa.