| Autor: |
Sarmientos, Paolo, Duchesne, Marc, Denèfle, Patrice, Boiziau, Janine, Fromage, Nadine, Delporte, Nadine, Parker, Fabienne, Lelièvre, Yves, Mayaux, Jean-François, Cartwright, Terence |
| Zdroj: |
Bio/Technology; May 1989, Vol. 7 Issue: 5 p495-501, 7p |
| Abstrakt: |
Using different types of efficient Escherichia coli expression systems, we have been able to obtain levels of human tissue-type plasminogen activator (tPA) reaching at least five percent of total bacterial proteins. The natural precursor of tPA (pre-tPA) expressed in E. coli accumulates, most probably in the cytoplasm, partly due to the low efficiency of the tPA leader sequence as a signal for secretion into the E. coli periplasm. We also expressed mature tPA in an insoluble, aggregated form and treatments were developed that permitted significant renaturation of this protein from the lysis pellet. Further characterization and purification of the rena-tured tPA polypeptide strongly suggest that a fully active enzyme, very similar to the eukaryotic product can be obtained, despite the total lack of glycosylation. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
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