Characterization of cell surface polypeptides of unfertilized, fertilized, and protease-treated zona-free mouse eggs.
The polypeptide composition of unfertilized, fertilized, and protease-treated zona-free mouse eggs was evaluated in this study. Zona-free eggs were radioiodinated by an Iodogen-catalyzed reaction. Light microscopic autoradiography of egg sections revealed that labeling was restricted to the cell surface. Labeled eggs were solubilized, and cell surface polypeptides were identified by one-dimensional SDS polyacrylamide gel electrophoresis and autoradiography. The unfertilized egg demonstrated 8-10 peptides that incorporated 125I, with major bands observed at approximately 145-150, 94, and 23 kilodaltons (kD). Zona-free eggs fertilized in vitro and then radiolabeled demonstrated several new bands in comparison to unfertilized eggs, with a major band appearing at approximately 36 kD. Treatment of radiolabeled unfertilized eggs with either trypsin or chymotrypsin (1 mg/ml for 5-20 min) caused enzyme-specific modifications in labeled polypeptides. Trypsin (T) treatment resulted in time-dependant modification of the three major peptides at 145-150, 94, and 23 kD. Chymotrypsin (CT) treatment, in contrast, was associated with loss or modification of the 94 kD band, with no apparent effect on either the 145-150 or 23 kD band. Taken together with previous data indicating that T or CT egg treatment interferes with sperm-egg attachment and fusion (Boldt et al.: Biol Reprod 39:19-27, 1988), these results suggest a possible role for the 94 kD protein in sperm-egg interaction.
Boldt
J
J
Department of Obstetrics/Gynecology, Medical College of Georgia, Augusta 30912.
Gunter
L E
LE
Howe
A M
AM
D000818:Animals; D001345:Autoradiography; D002462:Cell Membrane / Q000378:metabolism; D002918:Chymotrypsin; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005306:Fertilization; D007457:Iodine Radioisotopes; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D010063:Ovum / Q000378:metabolism*; D014238:Trichloroacetic Acid; D014357:Trypsin
D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D014238:Trichloroacetic Acid; D002918:Chymotrypsin; D014357:Trypsin