Direct characterization of factor VIII in plasma: Detection of a mutation altering a thrombin cleavage site (arginine-372 → histidine)

M. Arai, H. Inaba, M. Higuchi, S. E. Antonarakis, H. H. Kazazian, M. Fujimaki, L. W. Hoyer

Research output: Contribution to journalArticlepeer-review

55 Scopus citations

Abstract

An immunoadsorbent method has been developed for the direct analysis of normal and variant plasma factor VIII. Using this method, the molecular defect responsible for mild hemophilia A has been identified for a patient whose plasma factor VIII activity is 0.05 unit/ml, even though the factor VIII antigen content is 3.25 units/ml. Although the variant factor VIII has an apparently normal molecular mass and chain composition, the 92-kDa heavy chain accumulates when the variant protein is incubated with thrombin and the 44-kDa heavy chain fragment cannot be detected. In contrast, thrombin cleavage of the 80-kDa light chain to the 72-kDa fragment is normal. As these data indicate a loss of factor VIII cleavage by thrombin at arginine-372, the genetic defect was determined by polymerase-chain-reaction amplification of exon 8 of the factor VIII gene and direct sequencing of the amplified product. A single-base substitution (guanine → adenine) was identified that produces an arginine to histidine substitution at amino acid residue 372. These data identify the molecular basis of an abnormal factor VIII, 'factor VIII-Kumamoto', that lacks procoagulant function because of impaired thrombin activation.

Original languageEnglish (US)
Pages (from-to)4277-4281
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume86
Issue number11
DOIs
StatePublished - 1989
Externally publishedYes

ASJC Scopus subject areas

  • General

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