Expression and functional characterization of an abnormal platelet membrane glycoprotein Ibα (Met239 → Val) reported in patients with platelet-type yon Willebrand Disease

Takanori Moriki, Mitsuru Murata, Tetsuya Kitaguchi, Hironobu Anbo, Makoto Handa, Kiyoaki Watanabe, Hoyu Takahashi, Yasuo Ikeda

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Platelet-type von Willebrand disease (vWD) is a congenital bleeding disorder characterized by heightened ristocetin-induced platelet aggregation caused by abnormally high affinity between the platelet membrane glycoprotein (GP) Ib/IX complex and von Willebrand factor (vWF). Two distinct point mutations, Gly233 to Val and Met239 to Val, have been reported in GPIbα. We have constructed a recombinant GPIbα fragment containing the latter mutation, Met239 to Val (M239V) and characterized the mutant molecule using two methods, ie, interaction between soluble vWF and immobilized M239V and inhibition of platelet aggregation by purified soluble M239V. Spontaneous binding (ie, binding without any inducers) was observed between 125I-vWF and immobilized M239V but not between 125I-vWF and immobilized wild-type (WT) GPIbα. The addition of low concentrations of ristocetin (0.2 mg/mL) induced specific 125I-vWF binding to immobilized M239V, but not to WT GPIbα. At high concentrations of ristocetin (1.2 mg/mL), both WT GPIbα and M239V specifically bound to 125I-vWF. Thus, M239V reproduced the unique functional abnormality of the GPIb/IX complex in platelet-type vWD. Moreover, the purified soluble M239V inhibited platelet aggregation induced by low concentration of ristocetin (0.3 mg/mL) in platelet-rich plasma from a patient having Met238 to Val mutation, whereas purified WT did not. These results provide direct evidences that the reported point mutation is the responsible molecular basis of this disorder.

Original languageEnglish
Pages (from-to)698-705
Number of pages8
JournalBlood
Volume90
Issue number2
Publication statusPublished - 1997 Jul 15
Externally publishedYes

Fingerprint

Platelet Glycoprotein GPIb-IX Complex
von Willebrand Factor
Platelets
Ristocetin
Blood Platelets
Platelet Aggregation
Agglomeration
Point Mutation
Congenital, Hereditary, and Neonatal Diseases and Abnormalities
Mutation
Platelet-Rich Plasma
Hemorrhage
Plasmas
Molecules

ASJC Scopus subject areas

  • Hematology

Cite this

Expression and functional characterization of an abnormal platelet membrane glycoprotein Ibα (Met239 → Val) reported in patients with platelet-type yon Willebrand Disease. / Moriki, Takanori; Murata, Mitsuru; Kitaguchi, Tetsuya; Anbo, Hironobu; Handa, Makoto; Watanabe, Kiyoaki; Takahashi, Hoyu; Ikeda, Yasuo.

In: Blood, Vol. 90, No. 2, 15.07.1997, p. 698-705.

Research output: Contribution to journalArticle

Moriki, T, Murata, M, Kitaguchi, T, Anbo, H, Handa, M, Watanabe, K, Takahashi, H & Ikeda, Y 1997, 'Expression and functional characterization of an abnormal platelet membrane glycoprotein Ibα (Met239 → Val) reported in patients with platelet-type yon Willebrand Disease', Blood, vol. 90, no. 2, pp. 698-705.
Moriki, Takanori ; Murata, Mitsuru ; Kitaguchi, Tetsuya ; Anbo, Hironobu ; Handa, Makoto ; Watanabe, Kiyoaki ; Takahashi, Hoyu ; Ikeda, Yasuo. / Expression and functional characterization of an abnormal platelet membrane glycoprotein Ibα (Met239 → Val) reported in patients with platelet-type yon Willebrand Disease. In: Blood. 1997 ; Vol. 90, No. 2. pp. 698-705.
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abstract = "Platelet-type von Willebrand disease (vWD) is a congenital bleeding disorder characterized by heightened ristocetin-induced platelet aggregation caused by abnormally high affinity between the platelet membrane glycoprotein (GP) Ib/IX complex and von Willebrand factor (vWF). Two distinct point mutations, Gly233 to Val and Met239 to Val, have been reported in GPIbα. We have constructed a recombinant GPIbα fragment containing the latter mutation, Met239 to Val (M239V) and characterized the mutant molecule using two methods, ie, interaction between soluble vWF and immobilized M239V and inhibition of platelet aggregation by purified soluble M239V. Spontaneous binding (ie, binding without any inducers) was observed between 125I-vWF and immobilized M239V but not between 125I-vWF and immobilized wild-type (WT) GPIbα. The addition of low concentrations of ristocetin (0.2 mg/mL) induced specific 125I-vWF binding to immobilized M239V, but not to WT GPIbα. At high concentrations of ristocetin (1.2 mg/mL), both WT GPIbα and M239V specifically bound to 125I-vWF. Thus, M239V reproduced the unique functional abnormality of the GPIb/IX complex in platelet-type vWD. Moreover, the purified soluble M239V inhibited platelet aggregation induced by low concentration of ristocetin (0.3 mg/mL) in platelet-rich plasma from a patient having Met238 to Val mutation, whereas purified WT did not. These results provide direct evidences that the reported point mutation is the responsible molecular basis of this disorder.",
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AU - Anbo, Hironobu

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AU - Takahashi, Hoyu

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