Download PDF
Thromb Haemost 1998; 79(06): 1199-1207
DOI: 10.1055/s-0037-1615040
Rapid Communication
Schattauer GmbH

The cDNA Cloning and Molecular Characterization of a Snake Venom Platelet Glycoprotein Ib-binding Protein, Mamushigin, from Agkistrodon halys blomhoffii Venom[*]

Yoshihiko Sakurai
1   From the Departments of Pediatrics and Blood Transfusion Medicine, Nara Medical University, Kashihara, Nara
,
Yoshihiro Fujimura
1   From the Departments of Pediatrics and Blood Transfusion Medicine, Nara Medical University, Kashihara, Nara
,
Tetsuro Kokubo
2   Division of Gene Function in Animals, Nara Institute of Science and Technology, Takayama, Ikoma, Nara
,
Kouji Imamura
2   Division of Gene Function in Animals, Nara Institute of Science and Technology, Takayama, Ikoma, Nara
,
Tomihisa Kawasaki
,
Makoto Handa
4   Blood Transfusion Center, Keio University School of Medicine, Shinjyuku, Tokyo
,
Masami Suzuki
5   Institute for Comprehensive Medical Science, Fujita Health University School of Medicine, Toyoake, Aichi, Japan
,
Taei Matsui
5   Institute for Comprehensive Medical Science, Fujita Health University School of Medicine, Toyoake, Aichi, Japan
,
Koiti Titani
5   Institute for Comprehensive Medical Science, Fujita Health University School of Medicine, Toyoake, Aichi, Japan
,
Akira Yoshioka
1   From the Departments of Pediatrics and Blood Transfusion Medicine, Nara Medical University, Kashihara, Nara
› Author Affiliations
Further Information

Publication History

Received 24 September 1997

Accepted after revision 25 February 1998

Publication Date:
07 December 2017 (online)

    Permissions and Reprints

Summary

The entire cDNA sequences of a novel snake venom platelet glyco-protein (GP) Ib-binding protein (BP) composed of an α/β heterodimeric structure, termed mamushigin, from Agkistrodon halys blomhoffii were determined, that include the leader peptides (21/23 amino acid residues) and mature subunits (136/123 amino acid residues). The mature subunits of mamushigin are 37.5% identical, and showed a high degree of similarity (37.7-67.5% identity) with the respective subunits of group VII C-type lectins (19). The sequences of the leader peptides of the mamusigin subunits showed the highest similarity (α-73.9/ β-82.6%) with those of factor IX/X-BP from Trimeresurus flavoviridis, and the cleavage site residue in both proteins was the same Ala–1.

The GPIb-binding specificity of mamushigin is strongly supported by several lines of evidence, but mamushigin can directly aggregate normal platelets, similar to alboaggregin-B (AL-B) (1). This differs from other GPIb-BP’s. In mamushigin-treated platelets, serotonin was not released, and flow cytometric analysis using a monoclonal antibody PAC-1 totally excluded platelet GPIIb/IIIa activation. Mamushigin enhanced platelet aggregation at low-shear stress, and this effect totally disappeared in the presence of GPIb-receptor blockers specific for von Willebrand factor binding, but not by GPIIb/IIIa-receptor blockers. At high-shear stress, mamushigin blocked platelet aggregation in a dose-dependent manner, as seen with other GPIb-BP’s. This paper, therefore, describes the cDNA cloning and molecular characterization of mamushigin which has a different effect on platelet aggregation under different shear stress.

* This work was supported in part by a Research Grant (8A-1) for Cardiovascular Diseases to Yoshihiro Fujimura from the Japanese Ministry of Health and Welfare, and by Grants-in-Aid to Koiti Titani from the Fujita Health University and the Uehara Memorial Foundation.


 

  • References

  • 1 Peng M, Lu W, Kirby EP. Alboaggregin-B: A new platelet agonist that binds to platelet membrane glycoprotein Ib. Biochemistry 1991; 30: 11529-36.
    CrossrefPubMedGoogle Scholar
  • 2 Peng M, Lu W, Kirby EP. Characterization of three alboaggregins purified from Trimeresurus albolabris venom. Thromb Haemost 1992; 67: 702-7.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 3 Peng M, Lu W, Beviglia L, Niewiarowski S, Kirby EP. Echicetin: a snake venom protein that inhibits binding of von Willebrand factor and alboaggre-gins to platelet glycoprotein Ib. Blood 1993; 81: 2321-8.
    PubMedGoogle Scholar
  • 4 Chen Y-L, Tsai I-H. Functional and sequence characterization of agkicetin, a new glycoprotein Ib antagonist isolated from Agkistrodon acutus venom. Biochem Biophys Res Commun 1995; 210: 472-7.
    CrossrefPubMedGoogle Scholar
  • 5 Taniuchi Y, Kawasaki T, Fujimura Y, Suzuki M, Titani Y, Sakai Y, Kaku S, Hisamichi N, Satoh N, Takenaka T, Handa M, Sawai Y. Flavocetin-A and -B, two high molecular mass glycoprotein Ib binding proteins with high affinity purified from Trimeresurus flavoviridis venom, inhibit platelet aggregation at high shear stress. Biochem Biopys Acta 1995; 1244: 331-8.
    PubMedGoogle Scholar
  • 6 Fujimura Y, Ikeda Y, Miura S, Yoshida E, Shima H, Nishida S, Suzuki M, Titani K, Taniuchi Y, Kawasaki T. Isolation and characterization of jararaca GPIb-BP, a snake venom antagonist specific to platelet glyco-protein Ib. Thromb Haemost 1995; 74: 743-50.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Kawasaki T, Taniuchi Y, Hisamichi N, Fujimura Y, Suzuki M, Titani K, Sakai Y, Kaku S, Satoh N, Takenaka T, Handa M, Sawai Y. Tokaracetin, a new platelet antagonist that binds to platelet glycoprotein Ib and inhibits von Willebrand factor-dependent shear-induced platelet aggregation. Biochemical J 1995; 308: 947-53.
    CrossrefPubMedGoogle Scholar
  • 8 Andrews RK, Kroll MH, Ward CM, Rose JW, Scarborough RM, Smith AI, Lopez JA, Berndt MC. Binding of a novel 50-kilodalton alboaggregin from Trimeresurus albolabris and related viper venom proteins to the platelet membrane glycoprotein Ib-IX-V complex. Effect on platelet aggregation and glycoprotein Ib-mediated platelet activation. Biochemistry 1996; 35: 12629-39.
    CrossrefPubMedGoogle Scholar
  • 9 Ikeda Y, Handa M, Kawano K, Kamata T, Murata M, Araki Y, Anbo H, Kawai Y, Watanabe K, Itagaki I, Sakai K, Ruggeri ZM. The role of von Willebrand factor and fibrinogen in platelet aggregation under varying shear stress. J Clin Invest 1991; 87: 1234-40.
    CrossrefPubMedGoogle Scholar
  • 10 Moake JL, Turner NA, Stathopoulos NA, Nolasco L, Hellums JD. Shear-induced platelet aggregation can be mediated by vWF released from platelets, as well as exogenous large or unusually large vWF multimers, requires adenosine diphosphate, and is resistent to aspirin. Blood 1988; 71: 1366-74.
    PubMedGoogle Scholar
  • 11 Usami Y, Suzuki M, Yoshida E, Sakurai Y, Hirano K, Kawasaki T, Fujimura Y, Titani K. Primary structure of alboaggregin-B purified from the venom of Trimeresurus albolabris . Biochem Biophys Res Commun 1996; 219: 727-33.
    CrossrefPubMedGoogle Scholar
  • 12 Kawasaki T, Fujimura Y, Usami Y, Suzuki M, Miura S, Sakurai Y, Makita K, Taniuchi Y, Hirano K, Titani K. Complete amino acid sequence and identification of the platelet glycoprotein Ib-binding site of jararaca GPIbBP, a snake venom protein isolated from Bothrops jararaca . J Biol Chem 1996; 271: 10635-9.
    CrossrefPubMedGoogle Scholar
  • 13 Peng M, Holt JC, Niewiarowski S. Isolation, characterization and amino acid sequence of echicetin β subunit, a specific inhibitor of von Willebrand factor and thrombin interaction with glycoprotein Ib. Biochem Biophys Res Commun 1994; 205: 68-72.
    CrossrefPubMedGoogle Scholar
  • 14 Polgar J, Magnenat EM, Peitsch MC, Wells TNC, Saqi MSA, Clemetson KJ. Amino acid sequence of the α subunit and computer modelling of the α and β subunits of echicetin from the venom of Echis carinatus (saw-scaled viper). Biochem J 1997; 323: 533-7.
    CrossrefPubMedGoogle Scholar
  • 15 Read MS, Shermer RW, Brinkhous KM. Venom coagglutinin: an activator of platelet aggregation dependent on von Willebrand factor. Proc Natl Acad Sci USA 1978; 75: 4514-8.
    CrossrefPubMedGoogle Scholar
  • 16 Usami Y, Fujimura Y, Suzuki M, Ozeki Y, Nishio K, Fukui H, Titani K. Primary structure of two-chain botrocetin, a von Willebrand factor modulator purified from the venom of Bothrops jararaca . Proc Natl Acad Sci USA 1993; 90: 928-32.
    CrossrefPubMedGoogle Scholar
  • 17 Atoda H, Hyuga M, Morita T. The primary structure of coagulation factor IX/factor X-binding protein isolated from the venom of Trimeresurus flavoviridis . J Biol Chem 1991; 266: 14903-11.
    PubMedGoogle Scholar
  • 18 Hirabayashi J, Kusunoki T, Kasai K. Complete primary structure of a galactose-specific lectin from the venom of the rattlesnake Crotalus atrox . J Biol Chem 1991; 266: 2320-6.
    PubMedGoogle Scholar
  • 19 Drickamer K. Evolution of Ca2+-dependent animal lectins. Prog Nucleic Acid Res Mol Biol 1993; 45: 207-32.
    PubMedGoogle Scholar
  • 20 Matsuzaki R, Yoshihara E, Yamada M, Shima K, Atoda H, Morita T. cDNA cloning of IX/X-BP, a heterogeneous two-chain anticoagulant protein from snake venom. Biocem Biophy Res Commun 1996; 220: 382-7.
    PubMedGoogle Scholar
  • 21 De Marco L, Mazzucato M, Masotti A, Fenton JW II, Ruggeri ZM. Function of glycoprotein Ibα in platelet activation induced by α-thrombin. J Biol Chem 1991; 266: 23776-83.
    PubMedGoogle Scholar
  • 22 Hanson SR, Pareti FI, Ruggeri ZM, Marzec UM, Kunicki TJ, Montgomery RR, Zimmerman TS, Harker LA. Effects of monoclonal antibodies against the platelet glycoprotein IIb/IIIa complex on thrombosis and hemostasis in the baboon. J Clin Invest 1988; 81: 149-58.
    CrossrefPubMedGoogle Scholar
  • 23 Montgomery RR, Kunicki TJ, Taves C, Pidard D, Corcoran M. Diagnosis of Bernard-Soulier syndrome and Glanzmann’s thrombasthenia with a monoclonal assay on whole blood. J Clin Invest 1983; 71: 385-9.
    CrossrefPubMedGoogle Scholar
  • 24 Shattil SJ, Cunningham M, Hoxie JA. Detection of activated platelets in whole blood using activation-dependent monoclonal antibodies and flow cytometry. Blood 1987; 70: 307-15.
    PubMedGoogle Scholar
  • 25 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-5.
    CrossrefPubMedGoogle Scholar
  • 26 Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem 1987; 262: 10035-8.
    PubMedGoogle Scholar
  • 27 Pearson WR, Lipman DJ. Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 1988; 85: 2444-8.
    CrossrefPubMedGoogle Scholar
  • 28 Fraker PJ, Speck JC. Protein and cell membrane iodinations with sparingly soluble chloramide, 1,3,4,6,-tetrachloro-3a,6a-diphenylglycoluril. Biocem Biophys Res Commun 1978; 80: 849-57.
    PubMedGoogle Scholar
  • 29 von Heijne G. Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem 1983; 133: 17-21.
    CrossrefPubMedGoogle Scholar
  • 30 Nakai K, Kanehisa M. A knowledge base for predicting protein localization sites in eukaryotic cells. Genomics 1992; 14: 897-911.
    CrossrefPubMedGoogle Scholar
  • 31 Matsui T, Hamako J, Suzuki M, Hayashi N, Ito M, Makita K, Fujimura Y, Ozeki Y, Titani K. Complete amino acid sequence of bitiscetin, a novel von Willebrand factor modulator protein, purified from snake venom of Bitis arietans . Res Commun Cell Mol Biol 1997; 1: 271-84.
    PubMedGoogle Scholar