De novo design of alpha-amylase inhibitor: a small linear mimetic of macromolecular proteinaceous ligands
| Autor: | Manfred Pavlík, Martin Horn, Michael Mareš, Lucie Dolečková-Marešová |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Stereochemistry
Swine Clinical Biochemistry Amino Acid Motifs Inhibition kinetics Biology Ligands Biochemistry Molecular evolution Peptide Library Drug Discovery Animals Glycoside hydrolase Amylase Enzyme Inhibitors Molecular Biology Pharmacology Binding Sites Molecular Structure Molecular Mimicry Isothermal titration calorimetry General Medicine Fluorescence Kinetics biology.protein Molecular Medicine alpha-Amylases Binding Determinants Macromolecule |
| Zdroj: | Chemistrybiology. 12(12) |
| ISSN: | 1074-5521 |
| Popis: | We report a low molecular weight inhibitor of alpha-amylases based on a linear peptidic scaffold designed de novo through the use of combinatorial chemistry. The inhibitory motif denoted PAMI (peptide amylase inhibitor) was selected by using L-peptide libraries and was fine-tuned by the introduction of unnatural modifications. PAMI specifically inhibits glycoside hydrolases of family 13. Its interaction with porcine pancreatic alpha-amylase was characterized by inhibition kinetics, fluorescence competition assays with natural alpha-amylase inhibitors, and isothermal titration calorimetry. We demonstrate that the critical amino acid residues in PAMI are shared with those in the macromolecular proteinaceous inhibitors that, however, bind to alpha-amylases through a spatially scattered set of intermolecular contacts. Thus, natural molecular evolution as well as combinatorial evolution selected the same alpha-amylase binding determinants for completely different spatial frameworks. |
| Databáze: | OpenAIRE |
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