Purification of human brain tissue factor.
Tissue factor (factor III) is a lipoprotein cofactor which markedly enhances the catalytic effect of coagulation factor VIIa upon factors IX and X. Human tissue factor apoprotein was purified 53,000-fold to homogeneity from brain using acetone delipidation, Triton X-100 extraction, and affinity chromatography upon factor VII-agarose. The purified apoprotein has an apparent molecular weight of 44,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, an amino acid composition similar to bovine brain tissue factor, and an NH2-terminal amino acid sequence of Ser-X-Asn-Thr-Val-Ala-Val-Tyr-X-Tyr-X-Leu-Lys-(Ser)-Lys-Asn-Phe. Optimal relipidation of the tissue factor apoprotein was associated with a 5000-fold enhancement of clotting activity and occurred at a phospholipid/apoprotein (w/w) ratio of greater than 600.
D000596:Amino Acids / Q000032:analysis; D001059:Apoproteins / Q000302:isolation & purification; D001923:Brain Chemistry; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005169:Factor VIII / Q000302:isolation & purification; D006801:Humans; D008970:Molecular Weight; D010743:Phospholipids / Q000378:metabolism; D013925:Thromboplastin / Q000302:isolation & purification* / Q000378:metabolism
D000596:Amino Acids; D001059:Apoproteins; D010743:Phospholipids; D005169:Factor VIII; D013925:Thromboplastin