Skip to main content
Journal cover image

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

Publication ,  Journal Article
Ortel, TL; Moore, KD; Ezban, M; Kane, WH
Published in: Thromb Haemost
January 1996

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.

Duke Scholars

Published In

Thromb Haemost

ISSN

0340-6245

Publication Date

January 1996

Volume

75

Issue

1

Start / End Page

36 / 44

Location

Germany

Related Subject Headings

  • Recombinant Proteins
  • Recombinant Fusion Proteins
  • Protein Structure, Tertiary
  • Peptide Fragments
  • Molecular Sequence Data
  • Factor VIII
  • Factor V
  • Epitope Mapping
  • Cardiovascular System & Hematology
  • Amino Acid Sequence
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Ortel, T. L., Moore, K. D., Ezban, M., & Kane, W. H. (1996). Effect of heterologous factor V heavy chain sequences on the secretion of recombinant human factor VIII. Thromb Haemost, 75(1), 36–44.
Ortel, T. L., K. D. Moore, M. Ezban, and W. H. Kane. “Effect of heterologous factor V heavy chain sequences on the secretion of recombinant human factor VIII.Thromb Haemost 75, no. 1 (January 1996): 36–44.
Ortel TL, Moore KD, Ezban M, Kane WH. Effect of heterologous factor V heavy chain sequences on the secretion of recombinant human factor VIII. Thromb Haemost. 1996 Jan;75(1):36–44.
Ortel, T. L., et al. “Effect of heterologous factor V heavy chain sequences on the secretion of recombinant human factor VIII.Thromb Haemost, vol. 75, no. 1, Jan. 1996, pp. 36–44.
Ortel TL, Moore KD, Ezban M, Kane WH. Effect of heterologous factor V heavy chain sequences on the secretion of recombinant human factor VIII. Thromb Haemost. 1996 Jan;75(1):36–44.
Journal cover image

Published In

Thromb Haemost

ISSN

0340-6245

Publication Date

January 1996

Volume

75

Issue

1

Start / End Page

36 / 44

Location

Germany

Related Subject Headings

  • Recombinant Proteins
  • Recombinant Fusion Proteins
  • Protein Structure, Tertiary
  • Peptide Fragments
  • Molecular Sequence Data
  • Factor VIII
  • Factor V
  • Epitope Mapping
  • Cardiovascular System & Hematology
  • Amino Acid Sequence