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Purification and characterization of a vitelline coat lysin from Ciona intestinalis spermatozoa
Author(s) -
Marino Rita,
De Santis Rosaria,
Hirohashi Noritaka,
Hoshi Motonori,
Pinto Maria Rosaria,
Usui Noriko
Publication year - 1992
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.1080320412
Subject(s) - ciona intestinalis , biology , chymotrypsin , vitelline membrane , enzyme , size exclusion chromatography , lysozyme , polyacrylamide gel electrophoresis , biochemistry , lysin , sperm , molecular mass , chromatography , trypsin , escherichia coli , chemistry , microbiology and biotechnology , oocyte , embryo , botany , bacteriophage , gene
In Ciona intestinalis a chymotrypsin‐like activity is involved in sperm penetration of the egg vitelline coat. A chymotrypsin‐like enzyme has been purified from spermatozoa by a protocol including ion exchange chromatography, gel filtration, and native polyacrylamide gel electrophoresis. The purified enzyme resulted homogeneous when analyzed by SDS‐PAGE. The molecular weight of the chmotrypsin‐like enzyme was estimated to be 35 KDa by gel filtration and 24 KDa by SDS‐PAGE in nonreducing conditions. The pH optimum of the enzyme is 8.4 and its activity is enhanced by Ca 2+ . It shows the highest activity towards the synthetic substrate Suc‐Ala‐Ala‐Pro‐Phe‐AMC. Furthermore, by electron microscopy, the purified enzyme affects the structure of egg vitelline coat, and thus it fulfills one of the criteria of a lysin.
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