Purification and preliminary characterization of rabbit vitamin K-dependent coagulation proteins.
We describe herein a procedure for the purification of protein C, protein S, prothrombin, factor VII, and factor X to apparent homogeneity from rabbit plasma. The initial steps, which are common to the purification of vitamin K-dependent proteins from other mammalian species, include adsorption onto and elution from barium followed by anion exchange chromatography. Proteins were further purified using a variety of techniques, including affinity chromatography, gel filtration, and anion exchange chromatography in a Fast Protein Liquid Chromatography system. Significant structural homologies exist between rabbit, human, and bovine vitamin K-dependent proteins. As is true for protein C and factor X in human and bovine plasma, rabbit protein C and factor X are two-chain proteins which can be converted to active proteases by specific venom activators. Rabbit factor VII is also a two-chain protein and can restore coagulant activity to human or bovine plasma deficient in factor VII. In contrast, rabbit protein S and prothrombin are single chain proteins. In view of the well-described species specificity of many of the vitamin K-dependent proteins, purified rabbit coagulant and anticoagulant proteins should be useful in the development of animal models of coagulation and/or thrombosis.
Weinstein
R E
RE
Department of Medicine, University of Connecticut School of Medicine, Farmington.
Rickles
F R
FR
Walker
F J
FJ
D000818:Animals; D001777:Blood Coagulation / Q000502:physiology*; D001798:Blood Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D005167:Factor VII / Q000302:isolation & purification; D005170:Factor X / Q000302:isolation & purification; D006023:Glycoproteins / Q000302:isolation & purification; D011486:Protein C / Q000302:isolation & purification; D017293:Protein S; D011516:Prothrombin / Q000302:isolation & purification; D011817:Rabbits; D014812:Vitamin K / Q000502:physiology*
D001798:Blood Proteins; D006023:Glycoproteins; D011486:Protein C; D017293:Protein S; D014812:Vitamin K; D005167:Factor VII; D011516:Prothrombin; D005170:Factor X