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 chro...

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Bibliographic Details
Published in:The Journal of biological chemistry 1985-09, Vol.260 (20), p.10917-10920
Main Authors: Broze, G J, Leykam, J E, Schwartz, B D, Miletich, J P
Format: Article
Language:English
Subjects:
Amino Acids - analysis
Apoproteins - isolation & purification
Biological and medical sciences
Blood coagulation. Blood cells
Brain Chemistry
Chromatography, Affinity
Coagulation factors
Electrophoresis, Polyacrylamide Gel
Factor VIII - isolation & purification
Fundamental and applied biological sciences. Psychology
Humans
Molecular and cellular biology
Molecular Weight
Phospholipids - metabolism
Thromboplastin - isolation & purification
Thromboplastin - metabolism
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Summary: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.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(17)39121-4