Structural characterization of thyroglobulin type-1 domains of equistatin
| Autor: | Brigita Lenarčič, Vito Turk, Katja Galeša, Roger H. Pain, Maarten A. Jongsma |
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| Rok vydání: | 2003 |
| Předmět: |
Circular dichroism
Proteases Equistatin purification Biophysics Cathepsin D Biochemistry Thyroglobulin Pichia Domain conformation Pichia pastoris cysteine proteinases protease inhibitor chemistry.chemical_compound Structural Biology pichia-pastoris expression Papain Genetics medicine Cloning Molecular Molecular Biology Protein secondary structure Cathepsin biology Circular Dichroism Proteins secondary structure Cell Biology stability biology.organism_classification Protease inhibitor (biology) Recombinant Proteins Protein Structure Tertiary PRI Bioscience circular-dichroism Kinetics chemistry Thyroglobulin type-1 domain human cathepsin-d actinia-equina medicine.drug |
| Zdroj: | FEBS Letters 539 (2003) FEBS Letters, 539, 120-124 |
| ISSN: | 0014-5793 |
| Popis: | Equistatin is a protein composed of three thyroglobulin type-1 domains. It inhibits papain-like cysteine proteinases and the aspartic proteinase, cathepsin D. To determine the structural basis for this inhibition we cloned and expressed the separated domains (eq d-1, eq d-2, eq d-3) in Pichia pastoris. Kinetic constants for the interaction of eq d-1 with papain and that of eq d-2 with cathepsin D are of similar order (subnanomolar) and are comparable to the constants obtained for full-length equistatin. The target proteinase for the third domain remains unknown. Thus, we demonstrate here that thyroglobulin type-1 motifs per se are able to support specific structural features that enable them to inhibit proteases from different classes. The overall conformation of three domains in equistatin is such that the interaction of domains 1 or 2 with their respective target enzymes is not hindered sterically by either domain. In addition, we show that the interaction of eq d-2 with cathepsin D results in conformational changes, which is not the case for the eq d-1/papain interaction. |
| Databáze: | OpenAIRE |
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