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Purification and Characterization of the Egg Jelly Macromolecules, Sialoglycoprotein and Fucose Sulfate Glycoconjugate, of the Sea Urchin Hemicentrotus Pulcherrimus
Author(s) -
Shimizu Takeshi,
Kinoh Hiroaki,
Yamaguchi Masaaki,
Suzuki Norio
Publication year - 1990
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/j.1440-169x.1990.00473.x
Subject(s) - fucose , sialoglycoprotein , fucoidan , biochemistry , hemicentrotus , gel electrophoresis , chemistry , acrosome reaction , glycoconjugate , iodoacetic acid , polyacrylamide gel electrophoresis , biology , sea urchin , chromatography , sialic acid , glycoprotein , microbiology and biotechnology , polysaccharide , in vitro , enzyme
A sialoglycoprotein and a fucose sulfate glycoconjugate (FSG) were purified from egg jelly of the sea urchin Hemicentrotus pulcherrimus . Sialoglycoprotein which consisted of sialic acid (90%, w/w) and protein (10%, w/w) did not cause induction of the acrosome reaction and sperm isoagglutination. FSG which contained one mol sulfate/mol fucose possessed 2.0 times protein to fucose by weight. The proteins in intact FSG were separated to two major (258 kDa and 237 kDa) and one minor (120 kDa) proteins by SDS‐polyacrylamide gel electrophoresis (SDS‐PAGE) in the presence of 2‐mercaptoethanol (2‐ME) while the proteins could not be separated by HPLC in the presence of 0.1% SDS or SDS‐PAGE without 2‐ME. However, after carboxymethylation of FSG, two major (260 kDa and 240 kDa) proteins and two minor (140 kDa and 135 kDa) proteins were separated from the fucose sulfate moiety by HPLC in the presence of 0.1% SDS or SDS‐PAGE without 2‐ME. When FSG was first carboxymethylated with non‐radioactive iodoacetic acid and then reduced with 2‐ME and finally carboxymethylated with 14 C‐iodoacetic acid, the most of radioactivity was detected in 140 kDa and 135 kDa proteins. Carboxymethylted‐FSG was less potent than intact FSG in induction of the acrosome reaction. Fucoidan, a fucose sulfate polymer, did not induce the acrosome reaction.