Amino acid sequence and characterization of C-type lectin purified from the snake venom of Crotalus ruber

Jiharu Hamako, Yukiyo Suzuki, Nobuhiro Hayashi, Mina Kimura, Yasuhiro Ozeki, Keiichiro Hashimoto, Taei Matsui

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish
Pages (from-to)299-306
Number of pages8
JournalComparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
Volume146
Issue number3
DOIs
Publication statusPublished - 01-03-2007

Fingerprint

Crotalus
C-Type Lectins
Snake Venoms
Lectins
Amino Acid Sequence
Amino Acids
Venoms
Sugars
Galactosides
Platelets
Platelet Aggregation
Agglomeration
Affinity chromatography
Acetylgalactosamine
Divalent Cations
Hemagglutination
von Willebrand Factor
Amino Acid Substitution
Lactose
Electrophoresis

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Physiology
  • Molecular Biology

Cite this

Hamako, Jiharu ; Suzuki, Yukiyo ; Hayashi, Nobuhiro ; Kimura, Mina ; Ozeki, Yasuhiro ; Hashimoto, Keiichiro ; Matsui, Taei. / Amino acid sequence and characterization of C-type lectin purified from the snake venom of Crotalus ruber. In: Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology. 2007 ; Vol. 146, No. 3. pp. 299-306.
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Amino acid sequence and characterization of C-type lectin purified from the snake venom of Crotalus ruber. / Hamako, Jiharu; Suzuki, Yukiyo; Hayashi, Nobuhiro; Kimura, Mina; Ozeki, Yasuhiro; Hashimoto, Keiichiro; Matsui, Taei.

In: Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, Vol. 146, No. 3, 01.03.2007, p. 299-306.

Research output: Contribution to journalArticle

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