Methods and Compositions for Producing Active Vitamin K-Dependent Proteins

Abstract

The present invention provides a method of identifying a human subject having increased or decreased sensitivity to warfarin, comprising detecting in the subject the presence of a single nucleotide polymorphism in the VKOR gene, wherein the single nucleotide polymorphism is correlated with increased or decreased sensitivity to warfarin, thereby identifying the subject having increased or decreased sensitivity to warfarin.

1 . A method of increasing the amount of carboxylated vitamin K dependent protein in a cell, comprising introducing into a cell that expresses a first nucleic acid encoding a vitamin K dependent protein a second heterologous nucleic acid encoding vitamin K epoxide reductase (VKOR) under conditions whereby said first and second nucleic acids are expressed to produce a vitamin K dependent protein and VKOR, respectively. 
     
     
         2 . A method of increasing the carboxylation of a vitamin K dependent protein, comprising introducing into a cell that expresses a first nucleic acid encoding a vitamin K dependent protein a second heterologous nucleic acid encoding vitamin K epoxide reductase (VKOR) under conditions whereby said first and second nucleic acids are expressed to produce a vitamin K dependent protein and VKOR, respectively. 
     
     
         3 . A method of producing a carboxylated vitamin K dependent protein in a cell, comprising introducing into a cell that expresses a first nucleic acid encoding a vitamin K dependent protein a second heterologous nucleic acid encoding vitamin K epoxide reductase (VKOR) under conditions whereby said first and second nucleic acids are expressed to produce a vitamin K dependent protein and VKOR, respectively, wherein the amount of carboxylated vitamin K dependent protein produced in the cell in the presence of VKOR is increased as compared to the amount of carboxylated vitamin K dependent protein produced in the cell in the absence of VKOR. 
     
     
         4 . The method of  claim 1 , wherein the cell further comprises a third nucleic acid encoding a vitamin K dependent carboxylase. 
     
     
         5 . The method of  claim 1 , wherein said vitamin K dependent protein is selected from the group consisting of Factor VII, Factor IX, Factor X, Protein C, Protein S, prothrombin and any combination thereof. 
     
     
         6 . The method of  claim 1 , wherein said cell is a plant cell. 
     
     
         7 . The method of  claim 1 , wherein said cell is an insect cell. 
     
     
         8 . The method of  claim 1 , wherein said cell is an animal cell. 
     
     
         9 . The method of  claim 1 , wherein said vitamin K-dependent carboxylase is bovine vitamin K dependent carboxylase. 
     
     
         10 . The method of any of  claim 1 , wherein said vitamin K-dependent carboxylase is vitamin K gamma glutamyl carboxylase (VKGC).