Hemophilia A due to mutations that create new N-glycosylation sites

Ashraf Aly, Miyoko Higuchi, Carol K. Kasper, Haig H. Kazazian, Stylianos E. Antonarakis, Leon W. Hoyer

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

In studying the molecular defects responsible for cross-reacting material-positive hemophilia A, we have identified two patients in whom the nonfunctional factor VIII-like protein has abnormal, slower-moving heavy or light chains on SDS/PAGE. Both patients have severe hemophilia A (<1% of normal factor VIII activity) with a normal plasma level of factor VIII antigen. The molecular defects were identified by denaturing gradient gel electrophoresis screening of PCR-amplified products of the factor VIII-coding DNA sequence followed by nucleotide sequencing of the abnormal PCR products. In patient ARC-21, a methionine-to-threonine substitution at position 1772 in the factor VIII light chain creates a potential new N-glycosylation site at asparagine-1770. In patient ARC-22, an isoleucine-to-threonine substitution at position 566 creates a potential new N-glycosylation site at asparagine-564 in the A2 domain of the factor VIII heavy chain. The mobility of these chains on SDS/PAGE was normal after N-Glycanase digestion and procoagulant activity was generated - to a maximum of 23% and 45% of control normal plasma. Abnormal N-glycosylation, blocking factor VIII procoagulant activity, represents a newly recognized mechanism for the pathogenesis of severe hemophilia A.

Original languageEnglish (US)
Pages (from-to)4933-4937
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number11
StatePublished - 1992
Externally publishedYes

Fingerprint

Factor VIII
Hemophilia A
Glycosylation
Mutation
AIDS-Related Complex
Asparagine
Threonine
Polyacrylamide Gel Electrophoresis
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Light
Polymerase Chain Reaction
Denaturing Gradient Gel Electrophoresis
Isoleucine
varespladib methyl
Methionine
Digestion
Nucleotides
Antigens
Proteins

Keywords

  • Denaturing gradient gel electrophoresis
  • Factor VIII
  • N-glycanase

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Aly, A., Higuchi, M., Kasper, C. K., Kazazian, H. H., Antonarakis, S. E., & Hoyer, L. W. (1992). Hemophilia A due to mutations that create new N-glycosylation sites. Proceedings of the National Academy of Sciences of the United States of America, 89(11), 4933-4937.

Hemophilia A due to mutations that create new N-glycosylation sites. / Aly, Ashraf; Higuchi, Miyoko; Kasper, Carol K.; Kazazian, Haig H.; Antonarakis, Stylianos E.; Hoyer, Leon W.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 89, No. 11, 1992, p. 4933-4937.

Research output: Contribution to journalArticle

Aly, A, Higuchi, M, Kasper, CK, Kazazian, HH, Antonarakis, SE & Hoyer, LW 1992, 'Hemophilia A due to mutations that create new N-glycosylation sites', Proceedings of the National Academy of Sciences of the United States of America, vol. 89, no. 11, pp. 4933-4937.
Aly, Ashraf ; Higuchi, Miyoko ; Kasper, Carol K. ; Kazazian, Haig H. ; Antonarakis, Stylianos E. ; Hoyer, Leon W. / Hemophilia A due to mutations that create new N-glycosylation sites. In: Proceedings of the National Academy of Sciences of the United States of America. 1992 ; Vol. 89, No. 11. pp. 4933-4937.
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