Evidence for the occurrence of an ecto-(adenosine triphosphatase) in rat epididymal spermatozoa.
Intact spermatozoa from rat cauda epididymis possess a Mg2+-dependent ATPase activity that hydrolyses externally added [gamma-32P]ATP. The ATPase reaction was linear with time for approx. 6 min and there was no detectable uptake of ATP by these cells. The ATPase activity of the whole spermatozoa was not due to leakage of the intracellular enzymic activity, contamination of the broken cells or any possible cell damage during incubation and isolation of spermatozoa. The activity of the enzyme was strongly inhibited (approx. 85%) by p-chloromercuribenzenesulphonic acid (50 microM) or the diazonium salt of sulphanilic acid (50 microM), which are believed not to enter the cells, whereas ouabain (0.5 mM), NaF (10 mM), NaN3 (2.5 mM) and oligomycin (5 microM) had no appreciable effect on the activity of the spermatozoal APTase. There was little loss of ATPase activity from the cells when washed with 0.5 mM-EDTA and an iso-osmotic or hyperosmotic medium. These data are consistent with the view that the observed ATPase activity is located on the external surface of spermatozoa. The sperm ecto-ATPase activity is resistant to the action of proteinases (50 micrograms/ml), namely trypsin, chymotrypsin and Pronase. Studies with various unlabelled phosphate esters indicate that the sperm ecto-ATPase is not a non-specific phosphatase and it has high degree of substrate specificity for ATP.
D002731:4-Chloromercuribenzenesulfonate / Q000494:pharmacology; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000255:Adenosine Triphosphate; D000818:Animals; D003979:Diazonium Compounds / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004822:Epididymis; D006868:Hydrolysis; D066298:In Vitro Techniques; D008297:Male; D009943:Organophosphorus Compounds; D010447:Peptide Hydrolases; D051381:Rats; D013094:Spermatozoa / Q000187:drug effects / Q000201:enzymology*; D013379:Substrate Specificity; D013425:Sulfanilic Acids / Q000031:analogs & derivatives / Q000494:pharmacology
D003979:Diazonium Compounds; D009943:Organophosphorus Compounds; D013425:Sulfanilic Acids; D002731:4-Chloromercuribenzenesulfonate; D000255:Adenosine Triphosphate; D004492:Edetic Acid; D010447:Peptide Hydrolases; D000251:Adenosine Triphosphatases