Effect of heterologous factor V heavy chain sequences on the secretion of recombinant human factor VIII.

Published

Journal Article

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 VIII/factor V chimeras that contained heterologous sequences from the A1 and/or A2 domains. Substitution of the factor VIII A1 domain dramatically reduced secretion of factor V approximately 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 approximately 3-fold, whereas substitution of the factor V A2 domain actually reduced secretion approximately 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.

Full Text

Duke Authors

Cited Authors

  • Ortel, TL; Moore, KD; Ezban, M; Kane, WH

Published Date

  • January 1, 1996

Published In

Volume / Issue

  • 75 / 1

Start / End Page

  • 36 - 44

PubMed ID

  • 8713777

Pubmed Central ID

  • 8713777

Electronic International Standard Serial Number (EISSN)

  • 2567-689X

International Standard Serial Number (ISSN)

  • 0340-6245

Language

  • eng