Sperm protein (sp50) binds to acrosome and plasma membranes in a Ca(2+)-dependent manner: possible role in acrosome reaction.
Annexins are a family of Ca(2+)-binding proteins involved in the exocytotic process. The presence and the role of annexins in mammalian spermatozoa have not been well established. Two annexin-like proteins were obtained from guinea pig testis, a doublet of Mr 31-33 kD (p31/33) and a protein of Mr 50 kD (p50). Both proteins were able to bind to erythrocyte ghosts in a Ca(2+)-dependent fashion. Polyclonal antibodies against p31/33 reacted with two major proteins, Mrs 50 kD (sp50) and 42 kD (sp42), from mature and immature guinea pig spermatozoa. p50 and sp50 are likely the native proteins from testis and spermatozoa, respectively, and they are seemingly related. By immunofluorescence, sp50 was only found in the acrosome region of immature and capacitated and noncapacitated spermatozoa, and its location was intracellular. In spermatozoa undergoing acrosome reaction, sp50 was detected in the whole acrosome, while in spermatozoa that had undergone acrosome reaction sp50 was not detected. However, in the protein pattern of acrosome reaction vesicles, anti-p31/33 antibody revealed diffuse bands of Mr 35-38 kD. sp50 was able to bind to plasma membrane fragments and acrosome outer membrane from demembranated sperm in a Ca(2+)-dependent fashion. The presence of sp50 in the acrosome region, its distribution throughout the acrosome membrane just before the acrosome reaction, and its ability to bind both plasma and outer acrosome membranes in a Ca(2+)-dependent manner suggest that sp50 may participate in the acrosome reaction mechanism in guinea pig spermatozoa.
Othón Hernández
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Departamento de Biología Celular, Centro de Investigación y Estudios Avanzados del Instituto Politécnico Nacional, Mexico.
Roa-Espitia
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AL
Cruz Trejo
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J
Mújica
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A
D000177:Acrosome / Q000378:metabolism*; D000818:Animals; D017302:Annexins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology*; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000378:metabolism; D006168:Guinea Pigs; D006801:Humans; D008297:Male; D011817:Rabbits; D029589:Seminal Plasma Proteins; D013094:Spermatozoa / Q000737:chemistry / Q000276:immunology; D013737:Testis / Q000737:chemistry
D017302:Annexins; D002352:Carrier Proteins; D029589:Seminal Plasma Proteins; C425042:spermadhesin; D002118:Calcium