| Autor: |
Diaz-Collier JA; Monsanto Corporate Research, Chesterfield, MO 63198., Palmier MO, Kretzmer KK, Bishop BF, Combs RG, Obukowicz MG, Frazier RB, Bild GS, Joy WD, Hill SR, et. al. |
| Jazyk: |
angličtina |
| Zdroj: |
Thrombosis and haemostasis [Thromb Haemost] 1994 Mar; Vol. 71 (3), pp. 339-46. |
| Abstrakt: |
Human tissue factor pathway inhibitor (TFPI) was expressed in E. coli as a non-glycosylated protein with an additional alanine attached to the aminoterminus of the wild type molecule. High-level expression was obtained with pMON6875, a plasmid containing a tac promoter, Gene 10 leader from bacteriophage T7, methionine-alanine-TFPI coding sequence, and the p22 transcriptional terminator. In this system, TFPI accounted for about 5-10% of the total cell protein. The inclusion bodies containing. TFPI were sulfitolyzed, purified by anion-exchange chromatography, refolded through a disulfide interchange reaction, and further fractionated by Mono S cation exchange chromatography. The Mono S resin resolved a peak of highly active TFPI from relatively inactive and possibly misfolded molecules. The E. coli TFPI was shown to be about two-fold more active, on a molar basis, than full-length human SK hepatoma TFPI in a tissue factor-induced clotting assay in human plasma. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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