Substitution of Val for Met at residue 239 of platelet glycoprotein Ibα in Japanese patients with platelet-type von Willebrand disease

H. Takahashi, Mitsuru Murata, T. Moriki, H. Anbo, T. Furukawa, K. Nikkuni, A. Shibata, M. Handa, Y. Kawai, K. Watanabe, Y. Ikeda

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Abstract

Genomic DNA was studied from four patients with platelet-type yon Willebrand disease (vWD) from two Japanese families previously reported. The entire coding region of platelet glycoprotein (GP) Ibα, a component of the platelet receptor for von Willebrand factor (vWF), was examined by polymerase chain reaction (PCR) followed by direct DNA sequence analysis. A single point mutation was found in all patients resulting in substitution of Val (GTG) for Met (ATG) at residue 239 of GPIbα. All patients were heterozygous for the mutation, whereas none of the unaffected family members had an amino acid substitution at residue 239. Because the nucleotide substitution destroys an NIa III restriction site on GPIbα, PCR products were subjected to digestion with this enzyme; DNA fragments from both normal and mutant alleles were detected in all affected individuals. In allele-specific PCR, DNA was amplified from patients' genomic DNA using either adenine- or guanine- containing primers, whereas only adenine-containing primer successfully amplified DNA from normal individuals. Cloning of amplified DNA into bacteriophage M13mp19 and subsequent DNA sequence analysis confirmed the mutation in these families. The absence of the amino acid substitution at residue 239 of GPIbα in the normal individuals tested, together with the linkage of this substitution to the phenotypic expression of disease in these two families and in a family recently described suggest that this amino acid change is a molecular basis for platelet-type vWD, and the substitution may produce a quite similar phenotype to the one reported previously (Gly to Val at residue 233 of GPIbα).

Original languageEnglish
Pages (from-to)727-733
Number of pages7
JournalBlood
Volume85
Issue number3
Publication statusPublished - 1995

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Platelet Glycoprotein GPIb-IX Complex
Platelet Membrane Glycoproteins
Platelets
Substitution reactions
Polymerase chain reaction
DNA
Blood Platelets
DNA sequences
Adenine
Amino Acid Substitution
DNA Sequence Analysis
Amino Acids
Polymerase Chain Reaction
Alleles
Mutation
Bacteriophages
Cloning
Guanine
Reaction products
Point Mutation

ASJC Scopus subject areas

  • Hematology

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Takahashi, H., Murata, M., Moriki, T., Anbo, H., Furukawa, T., Nikkuni, K., ... Ikeda, Y. (1995). Substitution of Val for Met at residue 239 of platelet glycoprotein Ibα in Japanese patients with platelet-type von Willebrand disease. Blood, 85(3), 727-733.

Substitution of Val for Met at residue 239 of platelet glycoprotein Ibα in Japanese patients with platelet-type von Willebrand disease. / Takahashi, H.; Murata, Mitsuru; Moriki, T.; Anbo, H.; Furukawa, T.; Nikkuni, K.; Shibata, A.; Handa, M.; Kawai, Y.; Watanabe, K.; Ikeda, Y.

In: Blood, Vol. 85, No. 3, 1995, p. 727-733.

Research output: Contribution to journalArticle

Takahashi, H, Murata, M, Moriki, T, Anbo, H, Furukawa, T, Nikkuni, K, Shibata, A, Handa, M, Kawai, Y, Watanabe, K & Ikeda, Y 1995, 'Substitution of Val for Met at residue 239 of platelet glycoprotein Ibα in Japanese patients with platelet-type von Willebrand disease', Blood, vol. 85, no. 3, pp. 727-733.
Takahashi, H. ; Murata, Mitsuru ; Moriki, T. ; Anbo, H. ; Furukawa, T. ; Nikkuni, K. ; Shibata, A. ; Handa, M. ; Kawai, Y. ; Watanabe, K. ; Ikeda, Y. / Substitution of Val for Met at residue 239 of platelet glycoprotein Ibα in Japanese patients with platelet-type von Willebrand disease. In: Blood. 1995 ; Vol. 85, No. 3. pp. 727-733.
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abstract = "Genomic DNA was studied from four patients with platelet-type yon Willebrand disease (vWD) from two Japanese families previously reported. The entire coding region of platelet glycoprotein (GP) Ibα, a component of the platelet receptor for von Willebrand factor (vWF), was examined by polymerase chain reaction (PCR) followed by direct DNA sequence analysis. A single point mutation was found in all patients resulting in substitution of Val (GTG) for Met (ATG) at residue 239 of GPIbα. All patients were heterozygous for the mutation, whereas none of the unaffected family members had an amino acid substitution at residue 239. Because the nucleotide substitution destroys an NIa III restriction site on GPIbα, PCR products were subjected to digestion with this enzyme; DNA fragments from both normal and mutant alleles were detected in all affected individuals. In allele-specific PCR, DNA was amplified from patients' genomic DNA using either adenine- or guanine- containing primers, whereas only adenine-containing primer successfully amplified DNA from normal individuals. Cloning of amplified DNA into bacteriophage M13mp19 and subsequent DNA sequence analysis confirmed the mutation in these families. The absence of the amino acid substitution at residue 239 of GPIbα in the normal individuals tested, together with the linkage of this substitution to the phenotypic expression of disease in these two families and in a family recently described suggest that this amino acid change is a molecular basis for platelet-type vWD, and the substitution may produce a quite similar phenotype to the one reported previously (Gly to Val at residue 233 of GPIbα).",
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T1 - Substitution of Val for Met at residue 239 of platelet glycoprotein Ibα in Japanese patients with platelet-type von Willebrand disease

AU - Takahashi, H.

AU - Murata, Mitsuru

AU - Moriki, T.

AU - Anbo, H.

AU - Furukawa, T.

AU - Nikkuni, K.

AU - Shibata, A.

AU - Handa, M.

AU - Kawai, Y.

AU - Watanabe, K.

AU - Ikeda, Y.

PY - 1995

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