Harnessing the functional diversity of plant cystatins to design inhibitor variants highly active against herbivorous arthropod digestive proteases
| Autor: | Dominique Michaud, Charles Goulet, Jonathan Tremblay, Juan Vorster, Marie-Claire Goulet |
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| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
Proteases Phage display Protein family medicine.medical_treatment Mutagenesis (molecular biology technique) Cysteine Proteinase Inhibitors Biology urologic and male genital diseases 01 natural sciences Biochemistry 03 medical and health sciences medicine Animals Arthropods Molecular Biology reproductive and urinary physiology Plant Proteins 030304 developmental biology 2. Zero hunger chemistry.chemical_classification 0303 health sciences Protease Cell Biology Protein engineering Cystatins female genital diseases and pregnancy complications Amino acid Coleoptera Enzyme chemistry Docking (molecular) Cystatin Peptide Hydrolases 010606 plant biology & botany |
| Zdroj: | The FEBS Journal. 289:1827-1841 |
| ISSN: | 1742-4658 1742-464X |
| DOI: | 10.1111/febs.16288 |
| Popis: | Protein engineering approaches have been proposed to improve the inhibitory properties of plant cystatins against herbivorous arthropod digestive proteases, generally involving the site-directed mutagenesis of functionally relevant amino acids or the selection of improved inhibitor variants by phage display approaches. Here, we propose a novel approach where the function-related structural elements of a cystatin are substituted by the corresponding elements of an alternative cystatin. Inhibitory assays were first performed with 20 representative plant cystatins and model Cys proteases, including arthropod proteases, to appreciate the extent of functional variability among the plant cystatin family. The most, and less, potent of these cystatins were then used as 'donors' of structural elements to create hybrids of tomato cystatin SlCYS8 used as a model 'recipient' inhibitor. In brief, inhibitory activities against Cys proteases strongly differed from one plant cystatin to another, with Ki (papain) values diverging by more than 30-fold and inhibitory rates against arthropod proteases varying by up to 50-fold depending on the enzymes assessed. In line with theoretical assumptions from docking models generated for different Cys protease-cystatin combinations, structural element substitutions had a strong impact on the activity of recipient cystatin SlCYS8, positive or negative depending on the basic inhibitory potency of the donor cystatin. Our data confirm the wide variety of cystatin inhibitory profiles among plant taxa. They also demonstrate the usefulness of these proteins as a pool of discrete structural elements for the design of cystatin variants with improved potency against herbivorous pest digestive Cys proteases. |
| Databáze: | OpenAIRE |
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