Purification, primary structures and evolution of coagulant proteases from Deinagkistrodon actus venom

Nikolai N. Nikandrov, Masanobu Deshimaru, Ayako Tani, Takahito Chijiwa, Hiroki Shibata, Chang Chun Chang, Yasuyuki Fukumaki, Tatsumi Ito, Motonori Ohno

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Deinagkistrodon (formerly Agkistrodon) actus (Taiwan) snake venom was found to contain at least seven closely related coagulant proteases. One of them, named actibin, was purified to homogeneity by means of four chromatographic steps. Actibin acted on fibrinogen to form fibrin clots with extremely high specific activity of 1630 NIH units/mg and preferentially released fibrinopeptide A. Actibin was an acidic glycoprotein (pI 3.4) with molecular weight of 41,000, which was reduced to 28,800 after deglycosylation with N-glycanase. The k cat /K m values of actibin for hydrolysis of tosyl-l-arginine methyl ester and benzoyl-l-arginine p-nitroanilide were one-third to a half those for thrombin, reflecting a high potency of actibin in fibrinogen clotting. The amidase activities of actibin and its family proteases were inhibited by 3,4-dichloroisocoumarin, a serine protease inhibitor, indicating that actibin and its family proteases are serine proteases. Four cDNAs, named DaP1 and DaP7-DaP9, encoding D. actus coagulant proteases were cloned. All cDNAs contain an open reading frame of 780 bp coding for 260 amino acid residues, including a signal peptide of 24 amino acid residues. Their amino acid sequences predicted are highly homologous to one another with one to five amino acid substitutions. When four D. actus protease cDNAs were compared with the cDNAs coding for Trimeresurus flavoviridis and T. gramineus venom serine proteases, accelerated evolution was clearly observed. Similarity of the nucleotide sequences of four D. actus protease cDNAs with no synonymous and one to five nonsynonymous substitutions seems not to be in direct conformity with accelerated evolution. This possibly suggests that they have evolved to a similar direction to enhance their clotting activity rather than to produce other physiological activities.

Original languageEnglish
Pages (from-to)907-917
Number of pages11
JournalToxicon
Volume46
Issue number8
DOIs
Publication statusPublished - Dec 15 2005

Fingerprint

Coagulants
Venoms
Purification
Peptide Hydrolases
Complementary DNA
Amino Acids
Serine Proteases
amidase
Fibrinogen
Substitution reactions
Fibrinopeptide A
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Trimeresurus
Snake Venoms
Agkistrodon
Serine Proteinase Inhibitors
Protein Sorting Signals
Fibrin
Thrombin
Amino Acid Substitution

All Science Journal Classification (ASJC) codes

  • Toxicology

Cite this

Purification, primary structures and evolution of coagulant proteases from Deinagkistrodon actus venom. / Nikandrov, Nikolai N.; Deshimaru, Masanobu; Tani, Ayako; Chijiwa, Takahito; Shibata, Hiroki; Chang, Chang Chun; Fukumaki, Yasuyuki; Ito, Tatsumi; Ohno, Motonori.

In: Toxicon, Vol. 46, No. 8, 15.12.2005, p. 907-917.

Research output: Contribution to journalArticle

Nikandrov, NN, Deshimaru, M, Tani, A, Chijiwa, T, Shibata, H, Chang, CC, Fukumaki, Y, Ito, T & Ohno, M 2005, 'Purification, primary structures and evolution of coagulant proteases from Deinagkistrodon actus venom', Toxicon, vol. 46, no. 8, pp. 907-917. https://doi.org/10.1016/j.toxicon.2005.08.016
Nikandrov, Nikolai N. ; Deshimaru, Masanobu ; Tani, Ayako ; Chijiwa, Takahito ; Shibata, Hiroki ; Chang, Chang Chun ; Fukumaki, Yasuyuki ; Ito, Tatsumi ; Ohno, Motonori. / Purification, primary structures and evolution of coagulant proteases from Deinagkistrodon actus venom. In: Toxicon. 2005 ; Vol. 46, No. 8. pp. 907-917.
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