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Thromb Haemost 1996; 75(01): 036-044
DOI: 10.1055/s-0038-1650218
Original Article
Schattauer GmbH Stuttgart

Effect of Heterologous Factor V Heavy Chain Sequences on the Secretion of Recombinant Human Factor VIII

Thomas L Ortel
1   The Division of Hematology-Oncology, Departments of Medicine and Pathology, Duke University Medical Center, Durham, North Carolina, USA
,
Karen D Moore
1   The Division of Hematology-Oncology, Departments of Medicine and Pathology, Duke University Medical Center, Durham, North Carolina, USA
,
Mirella Ezban
2   The Division of Coagulation Research, Novo-Nordisk A/S, Copenhagen, Denmark
,
William H Kane
1   The Division of Hematology-Oncology, Departments of Medicine and Pathology, Duke University Medical Center, Durham, North Carolina, USA
› Author Affiliations
Further Information

Publication History

Received 02 August 1995

Accepted after revision 03 October 1995

Publication Date:
10 July 2018 (online)

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Summary

Factor VIII and factor V share a repetitive domain structure of A1-A2-B-A3-C1-C2. To define the region(s) within the factor VIII heavy chain that result in inefficient expression of the recombinant protein, we expressed a series of factor VIH/factor V chimeras that contained heterologous sequences from the A1 and/or A2 domains. Substitution of the factor VIIIA1 domain dramatically reduced secretion of factor V ~ 500-fold, whereas substitution of the factor VIII A2 domain had minimal effect on secretion. Conversely, substitution of the factor V A1 domain increased secretion of factor VIII ~3-fold, whereas substitution of the factor V A2 domain actually reduced secretion ~4-fold. Pulse chase experiments confirmed that reduced expression levels were due to decreased secretion rather than instability of secreted protein. Smaller substitutions did not further localize within the A1 domain the regions responsible for inefficient secretion.

 

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