Identification and characterization of membrane cofactor protein of human spermatozoa.
Membrane cofactor protein (MCP) regulates C activation by serving as a cofactor for the cleavage of C3b and C4b by the serine protease factor I. An MCP-like molecule on the inner acrosomal membrane of human spermatozoa has been characterized. Three mAb and a rabbit polyclonal antibody against MCP recognized the sperm protein. On SDS-PAGE, it migrated as a single band with a molecular mass of 38,000 and 44,000 Da under nonreducing or reducing conditions, respectively. The molecular mass was 10,000 to 20,000 Da less than the two forms of MCP expressed on others cells. The electrophoretic pattern, by one- and two-dimensional gel analysis, and the isoelectric point profile (4.5 to 5.0) of the sperm protein were similar among multiple individuals. In contrast to MCP of other cells, digestion with endoglycosidases did not alter either the m.w. or the pI of the protein, suggesting that it is a poorly or nonglycosylated form of MCP. The solubilized sperm protein bound C3 with broken thioester bond to Sepharose and possessed cofactor activity for factor I-mediated cleavage of C3 with the broken bond. A mAb that blocks the regulatory function of MCP inhibited the cofactor activity of the sperm lysate. Thus, the sperm protein is an antigenic and functional homologue of MCP but has the distinct structural features of a lower m.w. and an apparent lack of glycosylation. MCP may play an essential role in the survival of the acrosome-reacted spermatozoa by modulating C activation in the female genital tract.
Cervoni
F
F
Institut National de la Santé et de la Recherche Médicale U210, Faculté de Médecine, Nice, France.
Oglesby
T J
TJ
Adams
E M
EM
Milesifluet
C
C
Nickells
M
M
Fenichel
P
P
Atkinson
J P
JP
Hsi
B L
BL
D000177:Acrosome / Q000502:physiology; D015703:Antigens, CD; D002241:Carbohydrates / Q000032:analysis; D003167:Complement Activation; D003176:Complement C3 / Q000378:metabolism; D003165:Complement System Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D008297:Male; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D011233:Precipitin Tests; D013094:Spermatozoa / Q000737:chemistry*
D015703:Antigens, CD; C497857:CD46 protein, human; D002241:Carbohydrates; D003176:Complement C3; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D003165:Complement System Proteins