Mapping of the catalytic site of CHO-t-PA and the t-PA variant BM 06.022 by synthetic inhibitors and substrates
| Autor: | J. Stürzebecher, Stephan Fischer, U. Neumann, G.-B. Kresse, Ulrich Kohnert |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
medicine.medical_treatment
Molecular Sequence Data Peptide CHO Cells Transfection Cleavage (embryo) Biochemistry Substrate Specificity Cricetinae Escherichia coli medicine Animals Humans Trypsin Amino Acid Sequence Binding site Molecular Biology Peptide sequence chemistry.chemical_classification Oligopeptide Binding Sites Protease biology Thrombin Active site Recombinant Proteins Benzamidines Kinetics chemistry Tissue Plasminogen Activator Mutagenesis Site-Directed biology.protein Oligopeptides Research Article medicine.drug |
| Zdroj: | Protein Science. 1:1007-1013 |
| ISSN: | 1469-896X 0961-8368 |
| DOI: | 10.1002/pro.5560010806 |
| Popis: | BM 06.022 is a t-PA deletion variant that is produced as inactive inclusion bodies in Escherichia coli and transformed into the native form by an in vitro refolding process. Until now, no X-ray and NMR structures of BM 06.022 were available. Therefore a detailed kinetic analysis of the hydrolysis of peptide substrates and of the inhibition by several benzamidine-derived inhibitors was carried out in order to assess that the active site region of the protease domain of BM 06.022 is correctly structured in comparison with t-PA. Our data reveal that the single-chain as well as the two-chain form of BM 06.022 and native t-PA are similar in catalytic and in inhibitor binding properties. This indicates that the active site and the highly complex rearrangement of t-PA upon cleavage of the Arg275-Ile276 bond are maintained in BM 06.022. |
| Databáze: | OpenAIRE |
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