TY - JOUR
T1 - Amino acid sequence and characterization of C-type lectin purified from the snake venom of Crotalus ruber
AU - Hamako, Jiharu
AU - Suzuki, Yukiyo
AU - Hayashi, Nobuhiro
AU - Kimura, Mina
AU - Ozeki, Yasuhiro
AU - Hashimoto, Keiichiro
AU - Matsui, Taei
N1 - Funding Information:
We thank Dr. Koiti Titani, Fujita Health University, for helpful suggestions. We are grateful to Ms. Tomomi Matsuura, Ayako Hirai and Mr. Masami Suzuki at Fujita Health University for technical assistance. We thank Mr. Ronald G. Belisle for editing the manuscript. This work was supported by Grants-in-Aid from Japanese Ministry of Education Culture and Science (to T. M.) and Fujita Health University (to T. M. and J. H.).
PY - 2007/3
Y1 - 2007/3
N2 - Galactoside-binding lectin was purified from the snake venom of Crotalus ruber by affinity chromatography on a lactose-agarose column, and the complete amino acid sequence was determined. The C. ruber venom lectin (CRL) showed a single band of 28 kDa by SDS-polyacrylamide electrophoresis under non-reducing conditions, but it showed a single band of 15 kDa under reducing conditions, indicating that CRL is a disulfide-linked homodimer of 15 kDa subunit. CRL specifically recognized beta-galactosides such as thiodigalactoside followed by N-acetylgalactosamine when examined with their inhibitory effects on CRL-induced hemagglutination. A CRL subunit was composed of 135 residues containing nine Cys residues and showed a high similarity to other C-type galactoside-binding lectins from snake venoms. C. atrox lectin (CAL) showed almost the same sequence except for eight amino acid residues. Neither CRL nor CAL induced platelet aggregation by itself or inhibited platelet aggregation mediated by von Willebrand factor or fibrinogen with agonists. CRL showed a similar oligomeric form and the sugar specificity as CAL, but it showed different divalent cation sensitivity such as Mn2+ and Ni2+. Homology modeling suggested that the amino acid substitution found in CRL does not affect sugar recognition of the lectin but might alter the conformation and influence the sugar binding pocket induced by the metal-ion binding.
AB - Galactoside-binding lectin was purified from the snake venom of Crotalus ruber by affinity chromatography on a lactose-agarose column, and the complete amino acid sequence was determined. The C. ruber venom lectin (CRL) showed a single band of 28 kDa by SDS-polyacrylamide electrophoresis under non-reducing conditions, but it showed a single band of 15 kDa under reducing conditions, indicating that CRL is a disulfide-linked homodimer of 15 kDa subunit. CRL specifically recognized beta-galactosides such as thiodigalactoside followed by N-acetylgalactosamine when examined with their inhibitory effects on CRL-induced hemagglutination. A CRL subunit was composed of 135 residues containing nine Cys residues and showed a high similarity to other C-type galactoside-binding lectins from snake venoms. C. atrox lectin (CAL) showed almost the same sequence except for eight amino acid residues. Neither CRL nor CAL induced platelet aggregation by itself or inhibited platelet aggregation mediated by von Willebrand factor or fibrinogen with agonists. CRL showed a similar oligomeric form and the sugar specificity as CAL, but it showed different divalent cation sensitivity such as Mn2+ and Ni2+. Homology modeling suggested that the amino acid substitution found in CRL does not affect sugar recognition of the lectin but might alter the conformation and influence the sugar binding pocket induced by the metal-ion binding.
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U2 - 10.1016/j.cbpb.2006.11.022
DO - 10.1016/j.cbpb.2006.11.022
M3 - Article
C2 - 17251046
AN - SCOPUS:33847287225
SN - 1096-4959
VL - 146
SP - 299
EP - 306
JO - Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
JF - Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
IS - 3
ER -