The Sperm-surface glycoprotein, SGP, is necessary for fertilization in the frog, Xenopus laevis.
To identify a molecule involved in sperm-egg plasma membrane binding at fertilization, a monoclonal antibody against a sperm-surface glycoprotein (SGP) was obtained by immunizing mice with a sperm membrane fraction of the frog, Xenopus laevis, followed by screening of the culture supernatants based on their inhibitory activity against fertilization. The fertilization of both jellied and denuded eggs was effectively inhibited by pretreatment of sperm with intact anti-SGP antibody as well as its Fab fragment, indicating that the antibody recognizes a molecule on the sperm's surface that is necessary for fertilization. On Western blots, the anti-SGP antibody recognized large molecules, with molecular masses of 65-150 kDa and minor smaller molecules with masses of 20-28 kDa in the sperm membrane vesicles. SGP was distributed over nearly the entire surface of the sperm, probably as an integral membrane protein in close association with microfilaments. More membrane vesicles containing SGP bound to the surface were found in the animal hemisphere compared with the vegetal hemisphere in unfertilized eggs, but the vesicle-binding was not observed in fertilized eggs. These results indicate that SGP mediates sperm-egg membrane binding and is responsible for the establishment of fertilization in Xenopus.
Nagai
Keita
K
Laboratory of Molecular Developmental Biology, Graduate School of Medicine, Yamaguchi University, Yamaguchi 753-8512, Japan.
Ishida
Takuya
T
Hashimoto
Takafumi
T
Harada
Yuichirou
Y
Ueno
Shuichi
S
Ueda
Yasushi
Y
Kubo
Hideo
H
Iwao
Yasuhiro
Y
D000818:Animals; D015153:Blotting, Western; D005260:Female; D005306:Fertilization / Q000502:physiology*; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism / Q000502:physiology*; D010063:Ovum / Q000502:physiology; D013094:Spermatozoa / Q000378:metabolism* / Q000502:physiology*; D029867:Xenopus Proteins / Q000378:metabolism / Q000502:physiology*; D014982:Xenopus laevis / Q000378:metabolism* / Q000502:physiology*
D008562:Membrane Glycoproteins; D029867:Xenopus Proteins