Proteases released from Xenopus laevis eggs at activation and their role in envelope conversion.
During fertilization of the Xenopus laevis egg, the egg envelope is converted so that further sperm contact with the egg is prevented. In this study two envelope conversion reactions were investigated, envelope hardening and limited hydrolysis of two structurally related envelope glycoproteins. Both of these reactions were shown to be sensitive to protease inhibitors. In an attempt to identify egg proteases involved in envelope conversion, the medium around activated dejellied eggs was collected and analyzed. The exudate was able to convert isolated envelopes and, when the exudate was analyzed using peptide substrates, two major activities were found, one with a preference for cleavage after argininyl peptide bonds and one with a preference for phenylalaninyl peptide bonds. Analysis of exudate using SDS-polyacrylamide gel electrophoresis with gelatin cast into the gel showed two bands of proteolytic activity, one at Mr 45,000 that was identified as the trypsin-like activity and one at Mr 30,000 that was identified as the chymotrypsin-like activity. When cortical granule exocytosis was suppressed using ammonium chloride, release of the two exudate proteases was also suppressed. Studies of the envelope conversion reactions using protease inhibitors indicated that the chymotrysin-like protease was involved in envelope conversion once it had been activated by the trypsin-like protease.
Lindsay
L L
LL
Department of Biochemistry and Biophysics, University of California, Davis 95616.
Hedrick
J L
JL
D000818:Animals; D003594:Cytoplasmic Granules / Q000502:physiology; D005089:Exocytosis; D005260:Female; D006863:Hydrogen-Ion Concentration; D008297:Male; D008562:Membrane Glycoproteins / Q000502:physiology*; D008970:Molecular Weight; D009994:Osmolar Concentration; D010063:Ovum / Q000201:enzymology* / Q000502:physiology; D010447:Peptide Hydrolases / Q000378:metabolism* / Q000502:physiology; D011480:Protease Inhibitors / Q000494:pharmacology; D012995:Solubility; D013084:Sperm-Ovum Interactions; D013379:Substrate Specificity; D014817:Vitelline Membrane / Q000502:physiology; D014982:Xenopus laevis
D008562:Membrane Glycoproteins; D011480:Protease Inhibitors; D010447:Peptide Hydrolases