A mechanism for differential release of acrosomal enzymes during the acrosome reaction.
To study the organization of fertilization enzymes in the sperm acrosome, we isolated and characterized two physicochemically distinct acrosomal fractions of guinea-pig spermatozoa. A soluble fraction contained the 25,000-Mr acrosomal autoantigen, AA1, and most of the acrosomal hyaluronidase and dipeptidyl peptidase II activity. A particulate fraction, designated acrosomal matrix (AM), consisted of membraneless crescent-shaped structures, and contained most of the acrosomal proacrosin. The AM also contained a 28,000-Mr putative proacrosin-binding protein, and a very-high-Mr component which, on reduction, was dissociated into 48,000-Mr and 67,000-Mr subunits. Autoproteolytic dissolution of the AM correlated with proteolysis by acrosin of the 28,000-Mr and 48,000-Mr AM molecules. Components of both the AM and the soluble fraction were localized by immuno-electron microscopy to the electron-dense region of the guinea-pig sperm acrosome. We conclude that acrosomal molecules are segregated into soluble and matrix compartments. This segregation is a function of disulphide bonding and non-covalent interactions among the relatively few components of the AM. Association of acrosin with the AM may be the mechanism by which this enzyme's release from the spermatozoon during the acrosome reaction is delayed relative to the release of other acrosomal molecules.
Hardy
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DM
Department of Biochemistry and Biophysics, University of California, Davis 95616.
Oda
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MN
Friend
D S
DS
Huang
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TT
D000176:Acrosin / Q000378:metabolism*; D000177:Acrosome / Q000201:enzymology* / Q000502:physiology; D000818:Animals; D015153:Blotting, Western; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002458:Cell Fractionation; D004220:Disulfides / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000378:metabolism; D006168:Guinea Pigs; D008297:Male; D008854:Microscopy, Electron; D008970:Molecular Weight; D013094:Spermatozoa / Q000648:ultrastructure
D004220:Disulfides; D004792:Enzyme Precursors; D000001:Calcimycin; C022438:proacrosin; D000176:Acrosin; D002118:Calcium