Characterization of two antigenically related integral membrane proteins of the guinea pig sperm periacrosomal plasma membrane.
The periacrosomal plasma membrane of mammalian spermatozoa functions both in recognition and in binding of the egg's zona pellucida and in the acrosome reaction. This study characterizes two antigenically related proteins with molecular weights of 35 kD (PM35) and 52 kD (PM52) of the guinea pig sperm periacrosomal plasma membrane. Polyclonal antisera were prepared against electrophoretically purified PM35 or PM52. Each antiserum recognized both the 35-kD and 52-kD polypeptides on Western blots, indicating that they are structurally related. This conclusion was supported by peptide mapping experiments demonstrating comparably sized fragments of both PM35 and PM52. Both PM35 and PM52 behave as integral membrane proteins during phase-separation analysis with Triton X-114. Electron microscopic immunocytochemistry and differential fractionation of sperm membranes established that both PM35 and PM52 are exclusively localized to the periacrosomal plasma membrane. Three different antisera were used for ultrastructural studies, and each specifically bound the cytoplasmic but not the extracellular membrane surface. The electrophoretic mobilities of the PM35 and PM52 polypeptides were unchanged during sperm maturation and during the ionophore-induced acrosome reaction. The localization of PM35 and PM52 suggests a potential role for these integral plasma membrane proteins in signal transduction or membrane fusion events of the acrosome reaction.
Westbrook-Case
V A
VA
Department of Cell Biology, Vanderbilt University, Nashville, Tennessee 37232.
Winfrey
V P
VP
Olson
G E
GE
D000177:Acrosome / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D000939:Epitopes; D006168:Guinea Pigs; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D008970:Molecular Weight; D013079:Sperm Maturation; D013094:Spermatozoa / Q000378:metabolism* / Q000648:ultrastructure
D000939:Epitopes; D008565:Membrane Proteins