Enzymic characteristics of ecto-phosphoprotein phosphatase in goat epididymal intact spermatozoa.
Intact washed spermatozoa from goat cauda epididymis possess an ecto-phosphoprotein phosphatase that causes dephosphorylation of phosphoserine and phosphothreonine residues of exogenous 32P-labelled histones. The cell-bound ecto-enzyme has high affinity for proteins (histones, casein, phosvitin, and protamine) rather than phosphate esters, such as p-nitrophenyl phosphate, beta-glycerophosphate, AMP, and ATP. The activity of the enzyme is inhibited by 4 mM Mg2+, Ca2+, Mn2+, or Co2+. Pi (10 mM), NaF (10 mM), and Zn2+ (1 mM) inhibit the enzyme by approximately 50, 35, and 100%, respectively. Polyamines such as spermine and spermidine at 10 mM each caused significant inhibition (60 and 30%, respectively) of the cell-bound phosphoprotein phosphatase activity, whereas cAMP, orthovanadate, and calmodulin (with or without Ca2+) had no appreciable effect. Under the standard assay conditions, spermatozoa remain intact as evidenced by assay of cytosolic enzyme markers. Both the washed and "native" intact spermatozoa showed nearly the same specific activity of the ectoenzyme. The product of the reaction (Pi) was found in the extracellular medium. Sonication doubled the enzymic activity of the intact cells. The specific activity of the enzyme was nearly fourfold higher in the intact forwardly motile cells than the "composite" spermatozoa. These data provide further support for the localization of a phosphoprotein phosphatase on the external surface of spermatozoa and that the ectoenzyme may have a role in the regulation of flagellar motility.
Barua
M
M
Indian Institute of Chemical Biology, Calcutta.
Majumder
G C
GC
D000818:Animals; D002413:Cations, Divalent; D004822:Epididymis; D006041:Goats; D007700:Kinetics; D008297:Male; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010768:Phosphoserine; D010769:Phosphothreonine; D011073:Polyamines / Q000494:pharmacology; D013094:Spermatozoa / Q000201:enzymology*; D013379:Substrate Specificity
D002413:Cations, Divalent; D011073:Polyamines; D010769:Phosphothreonine; D010768:Phosphoserine; D010749:Phosphoprotein Phosphatases