Testicular biosynthesis and epididymal endoproteolytic processing of rat sperm surface antigen 2B1.
Binding of mammalian spermatozoa to the zona pellucida of homologous eggs is mediated by specific molecules on their surface membranes. In the present investigation we describe the biogenesis, epididymal processing and cellular distribution of a plasma membrane antigen (2B1) on rat spermatozoa that has a potential role in mediating zona binding. 2B1 is expressed postmeiotically in the testis as a precursor glycoprotein (approximately 60 kDa) that first appears on the plasma membrane of stage 6 to 8 round spermatids. Northern and western blot analyses show that there is a close correlation between the timing of transcription and expression of the glycoprotein on the cell surface. During spermatid elongation 2B1 is excluded from the head domain and is sequestered onto the sperm tail. As spermatozoa pass through the caput epididymidis 2B1 is endoproteolytically cleaved at a specific arginine residue (Arg 312) to produce a heterodimeric glycoprotein (approximately 40 kDa and approximately 19 kDa) containing intramolecular disulphide bridges. Endoproteolysis at Arg 312 also takes place during culture of washed testicular or caput spermatozoa in vitro and can be prevented by serine proteinase inhibitors or enhanced by trypsinisation. However, neither processing in vivo or in vitro has any effect on the domain organisation of 2B1 antigen i.e. it remains localised to the tail. These results support the hypothesis that sperm antigens that are important for fertilization are synthesized as precursor molecules in the testis and are then "activated' during epididymal maturation and capacitation, thereby ensuring that they only become fully functional at the site of fertilization.
Jones
R
R
Department of Signalling, Babraham Institute, Cambridge, UK.
Ma
A
A
Hou
S T
ST
Shalgi
R
R
Hall
L
L
D000818:Animals; D015153:Blotting, Western; D015815:Cell Adhesion Molecules; D011508:Chondroitin Sulfate Proteoglycans / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D010450:Endopeptidases / Q000494:pharmacology; D004822:Epididymis / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D006821:Hyaluronoglucosaminidase; D007150:Immunohistochemistry; D008297:Male; D011480:Protease Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D013087:Spermatids / Q000737:chemistry; D013090:Spermatocytes / Q000737:chemistry; D013091:Spermatogenesis; D013093:Spermatogonia / Q000737:chemistry; D013094:Spermatozoa / Q000737:chemistry*; D013737:Testis / Q000378:metabolism*
D015815:Cell Adhesion Molecules; D011508:Chondroitin Sulfate Proteoglycans; D011480:Protease Inhibitors; D012333:RNA, Messenger; D006821:Hyaluronoglucosaminidase; C066852:hyaluronidase PH-20; D010450:Endopeptidases