Regulation of proacrosin conversion in isolated guinea pig sperm acrosomal apical segments.
Following sodium dodecyl sulfate-polyacrylamide gel electrophoresis, proacrosin has been identified in extracts of intact guinea spermatozoa as a major silver staining band which reacted immunologically with antibodies made against purified proacrosin from guinea pig testis. Proacrosin exhibited an approximate Mr of 50,000 and was rapidly converted to an Mr 45,000 protein following induction of the acrosome reaction with 2.0 mM CaCl2 and 1 micrograms/ml A23187. Apical segments isolated at pH 6.0 from guinea pig spermatozoa also contained a major silver staining band of Mr 50,000 which cross-reacted with antibodies to guinea pig testis proacrosin. Subcellular fractionation of spermatozoa indicated that proacrosin remained in the particulate fraction of homogenized spermatozoa and was enriched within the isolated acrosomal apical segment. When apical segments isolated at pH 6.0 were incubated at pH 7.5, proacrosin was rapidly converted to the Mr 45,000 form observed in spermatozoa undergoing the acrosome reaction. The conversion process in isolated apical segments was inhibited by leupeptin and was accelerated in the presence of calcium, magnesium, and manganese. Zinc completely inhibited the conversion of proacrosin to the Mr 45,000 protein. Neither proacrosin nor the Mr 45,000 protein were released into the supernatant fluid during the incubation of apical segments at pH 7.5. Furthermore, the proteins were resistant to solubilization by 150 mM NaCl and 1% Triton X-100 but were solubilized by treatment of apical segments with 1 M NaCl. These results provide evidence as to the identity and subcellular distribution of proacrosin in intact guinea pig sperm prior to zymogen conversion and suggest that isolated apical segments exhibit a subset of the exocytotic reactions leading to completion of the acrosome reaction.
Noland
T D
TD
Department of Cell Biology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232.
Davis
L S
LS
Olson
G E
GE
D000176:Acrosin / Q000235:genetics* / Q000302:isolation & purification; D000177:Acrosome / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002462:Cell Membrane / Q000201:enzymology; D004792:Enzyme Precursors / Q000235:genetics* / Q000302:isolation & purification; D006168:Guinea Pigs; D008297:Male; D008970:Molecular Weight; D011499:Protein Processing, Post-Translational; D012697:Serine Endopeptidases / Q000235:genetics*; D013094:Spermatozoa / Q000201:enzymology*
D004792:Enzyme Precursors; D000001:Calcimycin; D012697:Serine Endopeptidases; C022438:proacrosin; D000176:Acrosin