The association of human coagulation factors VIII, IXa and X with phospholipid vesicles involves both electrostatic and hydrophobic interactions.
Blood coagulation factor X (FX) is converted to its active form (FXa) by a membrane bound multi-protein enzyme complex, comprised of factor VIII (FVIII), factor IXa (FIXa) and FX. Characterization of the molecular forces involved in the association of these proteins with phospholipids is crucial to understanding how these proteins bind to the lipid milieux of physiological membranes. In this report, the molecular forces involved in the association of FVIII, FIXa or FX with phospholipid vesicles (PLV) were characterized by ligand affinity chromatographic analyses. Treating FVIII-affinity columns with agents that disrupt electrostatic interactions caused elution of 15.2% of the total bound PLV, while agents that disrupt hydrophobic interactions caused elution of 84.8% of the total bound PLV. These results demonstrate that the association of PLV with FVIII is primarily hydrophobic. In contrast, the association of PLV with FIXa or FX is largely the result of electrostatic forces. This was established by observing that 71.3% and 78.9% of the total bound PLV was eluted from FIXa- and FX-affinity columns, respectively, by agents that disrupt electrostatic interactions. Of the total bound PLV, 28.7% and 21.2% were eluted from FIXa- and FX-affinity columns, respectively, by agents that disrupt hydrophobic interactions. These data demonstrate that hydrophobic forces play a heretofore unrecognized role in the association of PLV with FIXa or FX.
Atkins
J S
JS
Ottawa Blood Centre, Canadian Red Cross Blood Services.
Ganz
P R
PR
D001779:Blood Coagulation Factors / Q000737:chemistry* / Q000378:metabolism; D015949:Factor IXa / Q000737:chemistry / Q000378:metabolism; D005169:Factor VIII / Q000737:chemistry / Q000378:metabolism; D005170:Factor X / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances; D008566:Membranes / Q000737:chemistry; D009097:Multienzyme Complexes / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry* / Q000378:metabolism
D001779:Blood Coagulation Factors; D046911:Macromolecular Substances; D009097:Multienzyme Complexes; D010743:Phospholipids; D005169:Factor VIII; D005170:Factor X; D015949:Factor IXa