Isolation and Characterization of jararaca GPIb-BP, a Snake Venom Antagonist Specific to Platelet Glycoprotein lb

Y Fujimura; Y Ikeda; S Miura; E Yoshida; H Shima; S Nishida; M Suzuki; K Titani; Y Taniuchi; T Kawasaki

doi:10.1055/s-0038-1649807

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1995; 74(02): 743-750
DOI: 10.1055/s-0038-1649807

Original Article

Platelets

Schattauer GmbH Stuttgart

Isolation and Characterization of jararaca GPIb-BP, a Snake Venom Antagonist Specific to Platelet Glycoprotein lb^[*]

Y Fujimura

¹The Department of Blood Transfusion, Nara Medical University, Kashlhara, Nara, Japan

,

Y Ikeda

²The Department of Internal Medicine, Keio University School of Medicine, Shinjuku, Tokyo, Japan

,

S Miura

¹The Department of Blood Transfusion, Nara Medical University, Kashlhara, Nara, Japan

,

E Yoshida

¹The Department of Blood Transfusion, Nara Medical University, Kashlhara, Nara, Japan

,

H Shima

¹The Department of Blood Transfusion, Nara Medical University, Kashlhara, Nara, Japan

,

S Nishida

¹The Department of Blood Transfusion, Nara Medical University, Kashlhara, Nara, Japan

,

M Suzuki

³The Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi, Japan

,

K Titani

³The Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi, Japan

,

Y Taniuchi

⁴The Research Institute of Yamanouchi Pharmaceutical Co., Ibaraki, Japan

,

T Kawasaki

⁴The Research Institute of Yamanouchi Pharmaceutical Co., Ibaraki, Japan

› Author Affiliations

Abstract

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Summary

A platelet glycoprotein lb-binding protein (GPIb-BP) was isolated from the snake venom of Bothrops jararaca. Jararaca GPIb-BP showed a single band with M_r of 30,000, and two distinct bands with M_r. of 17,000/13,000 under non-reducing and reducing conditions, respectively, on SDS-polyacrylamide gel electrophoresis. Jararaca GPIb-BP itself induced neither platelet aggregation nor serotonin release from platelets, but specifically bound to GPIb (40,629 ± 2,521 molecules per normal platelet, with Kd 39.1 ± 2.4 nM at saturation). The purified venom protein completely inhibited ristocetin- or botrocetin-induccd von Willebrand factor (vWF) binding, and blocked the bovine vWF binding to GPIb, with IC₅₀ values ranging from 28 to 42 nM, without affecting the platelet aggregation induced by ADP or α-thrombin. ¹²⁵1-jararaca GPIb-BP binding to GPIb was not altered by the presence of human α-thrombin. Jararaca GPIb-BP at a final concentration of 104 nM totally abolished vWF-dependent shear- induced platelet aggregation (SIPA) at a high shear stress, but had no effect on SIPA at a low shear stress. Reduced and S-carboxyamidomethylated jararaca GPIb-BP lost its inhibitory activity on SIPA. The NH²-terminal amino acid sequences of the subunits revealed a high degree of homology with those of several Ca²⁺-dependent lectins, especially to those of two functionally opposite venom proteins, botrocetin (a vWF-modulator) and alboaggregin-B (a GPIb- modulator).

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References

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Isolation and Characterization of jararaca GPIb-BP, a Snake Venom Antagonist Specific to Platelet Glycoprotein lb[*]

Isolation and Characterization of jararaca GPIb-BP, a Snake Venom Antagonist Specific to Platelet Glycoprotein lb^[*]