Intracellular accumulation of factor VIII induced by missense mutations Arg593-->Cys and Asn618-->Ser explains cross-reacting material-reduced haemophilia A.

Cys and Asn618-->Ser explains cross-reacting material-reduced haemophilia A. -->
Autoři: Roelse JC; Department of Blood Coagulation, CLB, Amsterdam, The Netherlands., De Laaf RT, Timmermans SM, Peters M, Van Mourik JA, Voorberg J
Zdroj: British journal of haematology [Br J Haematol] 2000 Feb; Vol. 108 (2), pp. 241-6.
Způsob vydávání: Journal Article
Jazyk: English
Informace o časopise: Publisher: Wiley-Blackwell Country of Publication: England NLM ID: 0372544 Publication Model: Print Cited Medium: Print ISSN: 0007-1048 (Print) Linking ISSN: 00071048 NLM ISO Abbreviation: Br J Haematol Subsets: MEDLINE
Imprint Name(s): Publication: Oxford : Wiley-Blackwell
Original Publication: Oxford : Blackwell Scientific Publications
Výrazy ze slovníku MeSH: Factor VIII/*genetics , Hemophilia A/*genetics , Introns/*genetics , Mutation, Missense/*genetics, Cross Reactions ; Factor VIII/metabolism ; Humans
Abstrakt: Patients with cross-reacting material (CRM)-reduced haemophilia A exhibit reduced levels of factor VIII antigen. In this study, we determined the molecular basis of the genetic defect in the factor VIII gene induced by either the Arg593-->Cys or the Asn618-->Ser missense mutation, identified in two CRM-reduced haemophilia A patients. We introduced either the Arg593-->Cys or the Asn618-->Ser mutation into a B-domain-deleted factor VIII cDNA and expressed the modified cDNAs in C127 cells. Reduced levels of factor VIII activity and factor VIII antigen in conditioned medium of transfected cells indicated that the secretion of both factor VIII variants was impaired. The ratio of factor VIII antigen present in cell extract to that in conditioned medium was 1.9 and 2.4 times higher for rFVIII-R593C and rFVIII-N618S, respectively, than for rFVIII. Metabolic labelling and immunoprecipitation of transfected cells revealed that rFVIII-R593C and rFVIII-N618S persisted somewhat longer inside the cell than factor rFVIII. Intracellular accumulation and subsequent degradation of factor VIII-R593C and factor VIII-N618S may explain the reduced levels of both factor VIII activity and antigen in plasma of mild haemophilia A patients with corresponding genetic defects.
Substance Nomenclature: 9001-27-8 (Factor VIII)
Entry Date(s): Date Created: 20000226 Date Completed: 20000413 Latest Revision: 20190705
Update Code: 20240829
DOI: 10.1046/j.1365-2141.2000.01834.x
PMID: 10691849
Autor: Roelse JC; Department of Blood Coagulation, CLB, Amsterdam, The Netherlands., De Laaf RT, Timmermans SM, Peters M, Van Mourik JA, Voorberg J
Jazyk: angličtina
Zdroj: British journal of haematology [Br J Haematol] 2000 Feb; Vol. 108 (2), pp. 241-6.
DOI: 10.1046/j.1365-2141.2000.01834.x
Abstrakt: Patients with cross-reacting material (CRM)-reduced haemophilia A exhibit reduced levels of factor VIII antigen. In this study, we determined the molecular basis of the genetic defect in the factor VIII gene induced by either the Arg593-->Cys or the Asn618-->Ser missense mutation, identified in two CRM-reduced haemophilia A patients. We introduced either the Arg593-->Cys or the Asn618-->Ser mutation into a B-domain-deleted factor VIII cDNA and expressed the modified cDNAs in C127 cells. Reduced levels of factor VIII activity and factor VIII antigen in conditioned medium of transfected cells indicated that the secretion of both factor VIII variants was impaired. The ratio of factor VIII antigen present in cell extract to that in conditioned medium was 1.9 and 2.4 times higher for rFVIII-R593C and rFVIII-N618S, respectively, than for rFVIII. Metabolic labelling and immunoprecipitation of transfected cells revealed that rFVIII-R593C and rFVIII-N618S persisted somewhat longer inside the cell than factor rFVIII. Intracellular accumulation and subsequent degradation of factor VIII-R593C and factor VIII-N618S may explain the reduced levels of both factor VIII activity and antigen in plasma of mild haemophilia A patients with corresponding genetic defects.
Databáze: MEDLINE
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