Skip to main content
construction release_alert
Scholars@Duke will be undergoing maintenance April 11-15. Some features may be unavailable during this time.
cancel
Journal cover image

Contributions of Asn2198, Met2199, and Phe2200 in the factor VIII C2 domain to cofactor activity, phospholipid-binding, and von Willebrand factor-binding.

Publication ,  Journal Article
Lewis, DA; Pound, ML; Ortel, TL
Published in: Thromb Haemost
May 2003

The crystal structure of the factor VIII C2 domain consists of a beta-sandwich core from which beta-hairpins and loops extend to form a hydrophobic surface. The hydrophobic surface includes M2199 and F2200 at the tip of the 1(st) beta-hairpin. To determine the individual contributions of residues N2198, M2199, and F2200 to phospholipid and von Willebrand factor (vWF) binding properties of factor VIII, we prepared mutant proteins with single alanine substitutions. We found that single mutations at N2198 and M2199 had relatively little impact on cofactor activity, or phospholipid and vWF binding. However the F2200A mutant had slightly lower cofactor activity at subsaturating phospholipid concentrations. Competitive ELISAs suggested that F2200 plays a more important role in both phospholipid-binding and vWF-binding than N2198 and M2199. All mutant proteins were still recognized by a monoclonal antibody and two factor VIII inhibitors that neutralized cofactor activity and blocked factor VIII binding to phospholipids.

Duke Scholars

Published In

Thromb Haemost

ISSN

0340-6245

Publication Date

May 2003

Volume

89

Issue

5

Start / End Page

795 / 802

Location

Germany

Related Subject Headings

  • von Willebrand Factor
  • Protein Structure, Tertiary
  • Protein Binding
  • Phospholipids
  • Phosphatidylserines
  • Mutation, Missense
  • Mutagenesis, Site-Directed
  • Liposomes
  • Humans
  • Factor VIII
 
Journal cover image

Published In

Thromb Haemost

ISSN

0340-6245

Publication Date

May 2003

Volume

89

Issue

5

Start / End Page

795 / 802

Location

Germany

Related Subject Headings

  • von Willebrand Factor
  • Protein Structure, Tertiary
  • Protein Binding
  • Phospholipids
  • Phosphatidylserines
  • Mutation, Missense
  • Mutagenesis, Site-Directed
  • Liposomes
  • Humans
  • Factor VIII