The Recombinant Catalytic Domain of Human Neutrophil Collagenase Lacks Type I Collagen Substrate Specificity
| Autor: | T. Kleine, A. Hillemann, Harald Tschesche, Susanne Schnierer, T. Gote, V Knäuper |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Neutrophils
Molecular Sequence Data Biophysics medicine.disease_cause Polymerase Chain Reaction Biochemistry Substrate Specificity law.invention chemistry.chemical_compound Affinity chromatography law Escherichia coli medicine Humans Amino Acid Sequence Collagenases Molecular Biology chemistry.chemical_classification Enzyme Precursors Base Sequence Hemopexin DNA Cell Biology Recombinant Proteins Enzyme chemistry Recombinant DNA Collagenase Collagen Type I collagen medicine.drug |
| Zdroj: | Biochemical and Biophysical Research Communications. 191:319-326 |
| ISSN: | 0006-291X |
| Popis: | The coding region for human neutrophil short form procollagenase lacking the hemopexin like domain coding region was amplified by polymerase chain reaction. Recombinant short form procollagenase was expressed in E. coli and purified in a three step procedure. Renaturation of this proenzyme was carried out by an effective new method using Q-Sepharose chromatography. Treatment of short form procollagenase with mercurials resulted in active short form collagenase M(r) 21,000 and an intermediate product of M(r) 23,000. These two products were separated by hydroxamate affinity chromatography. The active, short form collagenase M(r) 21,000 is stable. Despite full proteolytic activity, it lacks type I collagen substrate specificity and forms the basis for crystallisation experiments. |
| Databáze: | OpenAIRE |
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