Manipulation of the membrane binding site of vitamin K-dependent proteins: enhanced biological function of human factor VII.
Recent studies suggested that modification of the membrane contact site of vitamin K-dependent proteins may enhance the membrane affinity and function of members of this protein family. The properties of a factor VII mutant, factor VII-Q10E32, relative to wild-type factor VII (VII, containing P10K32), have been compared. Membrane affinity of VII-Q10E32 was about 20-fold higher than that of wild-type factor VII. The rate of autoactivation VII-Q10E32 with soluble tissue factor was 100-fold faster than wild-type VII and its rate of activation by factor Xa was 30 times greater than that of wild-type factor VII. When combined with soluble tissue factor and phospholipid, activated factor VII-Q10E32 displayed increased activation of factor X. Its coagulant activity was enhanced in all types of plasma and with all sources of tissue factor tested. This difference in activity (maximum 50-fold) was greatest when coagulation conditions were minimal, such as limiting levels of tissue factor and/or phospholipid. Because of its enhanced activity, factor VII-Q10E32 and its derivatives may provide important reagents for research and may be more effective in treatment of bleeding and/or clotting disorders.
Shah
A M
AM
Department of Biochemistry, University of Minnesota, 1479 Gortner Avenue, St. Paul, MN 55108, USA.
Kisiel
W
W
Foster
D C
DC
Nelsestuen
G L
GL
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D005167:Factor VII / Q000737:chemistry* / Q000378:metabolism*; D005170:Factor X / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D010743:Phospholipids / Q000494:pharmacology; D017354:Point Mutation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013925:Thromboplastin / Q000378:metabolism; D014162:Transfection; D014812:Vitamin K / Q000378:metabolism*
D010446:Peptide Fragments; D010743:Phospholipids; D011994:Recombinant Proteins; D014812:Vitamin K; D005167:Factor VII; D005170:Factor X; D013925:Thromboplastin