Factor VII and protein C are phosphatidic acid-binding proteins

Biochemistry. 2013 Aug 20;52(33):5545-52. doi: 10.1021/bi4006368. Epub 2013 Aug 7.


Seven proteins in the human blood clotting cascade bind, via their GLA (γ-carboxyglutamate-rich) domains, to membranes containing exposed phosphatidylserine (PS), although with membrane binding affinities that vary by 3 orders of magnitude. Here we employed nanodiscs of defined phospholipid composition to quantify the phospholipid binding specificities of these seven clotting proteins. All bound preferentially to nanobilayers in which PS headgroups contained l-serine versus d-serine. Surprisingly, however, nanobilayers containing phosphatidic acid (PA) bound substantially more of two of these proteins, factor VIIa and activated protein C, than did equivalent bilayers containing PS. Consistent with this finding, liposomes containing PA supported higher proteolytic activity by factor VIIa and activated protein C toward their natural substrates (factors X and Va, respectively) than did PS-containing liposomes. Moreover, treating activated human platelets with phospholipase D enhanced the rates of factor X activation by factor VIIa in the presence of soluble tissue factor. We hypothesize that factor VII and protein C bind preferentially to the monoester phosphate of PA because of its accessibility and higher negative charge compared with the diester phosphates of most other phospholipids. We further found that phosphatidylinositol 4-phosphate, which contains a monoester phosphate attached to its myo-inositol headgroup, also supported enhanced enzymatic activity of factor VIIa and activated protein C. We conclude that factor VII and protein C bind preferentially to monoester phosphates, which may have implications for the function of these proteases in vivo.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • 1-Carboxyglutamic Acid / chemistry
  • 1-Carboxyglutamic Acid / metabolism*
  • Binding Sites
  • Binding, Competitive
  • Factor VII / chemistry
  • Factor VII / metabolism*
  • Humans
  • Kinetics
  • Lipid Bilayers / chemistry
  • Lipid Bilayers / metabolism
  • Liposomes / chemistry
  • Liposomes / metabolism
  • Models, Molecular
  • Phosphatidic Acids / chemistry
  • Phosphatidic Acids / metabolism*
  • Phosphatidylinositol Phosphates / chemistry
  • Phosphatidylinositol Phosphates / metabolism
  • Phosphatidylserines / chemistry
  • Phosphatidylserines / metabolism
  • Protein Binding
  • Protein C / chemistry
  • Protein C / metabolism*
  • Surface Plasmon Resonance


  • Lipid Bilayers
  • Liposomes
  • Phosphatidic Acids
  • Phosphatidylinositol Phosphates
  • Phosphatidylserines
  • Protein C
  • phosphatidylinositol 4-phosphate
  • 1-Carboxyglutamic Acid
  • Factor VII