Multiple pathways guide oxygen diffusion into flavoenzyme active sites
| Autor: | Pimchai Chaiyen, Marco W. Fraaije, Willem J. H. van Berkel, Pirom Chenprakhon, J. Andrew McCammon, Riccardo Baron, Andrea Mattevi, Conor T. Riley, Federico Forneris, Andrea Alfieri, Remko T. Winter, Kittisak Thotsaporn |
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| Přispěvatelé: | Biotechnology |
| Jazyk: | angličtina |
| Rok vydání: | 2009 |
| Předmět: |
Models
Molecular Cholesterol oxidase molecular-dynamics Crystallography X-Ray 01 natural sciences Biochemistry Diffusion Molecular dynamics Catalytic Domain 0303 health sciences Oxidase test Multidisciplinary biology Chemistry computational biochemistry flavin Physical Sciences fluorescence simulations Oxidoreductases Protein Binding Stereochemistry enzymology Flavoprotein Biochemie Streptomyces coelicolor Flavin group 010402 general chemistry Catalysis Cofactor o-2 Enzyme catalysis 03 medical and health sciences Flavins cholesterol oxidase Computer Simulation oxygen reactivity x-ray crystallography 030304 developmental biology VLAG Flavoproteins Monooxygenase streptomyces-coelicolor Protein Structure Tertiary 0104 chemical sciences Oxygen Alcohol Oxidoreductases enzyme Mutagenesis Site-Directed biology.protein alditol oxidase protein |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America, 106(26), 10603-10608 Proceedings of the National Academy of Sciences of the United States of America 106 (2009) 26 Proceedings of the National Academy of Sciences of the United States of America, 106(26), 10603-10608. NATL ACAD SCIENCES |
| ISSN: | 0027-8424 |
| Popis: | Dioxygen (O 2 ) and other gas molecules have a fundamental role in a variety of enzymatic reactions. However, it is only poorly understood which O 2 uptake mechanism enzymes employ to promote efficient catalysis and how general this is. We investigated O 2 diffusion pathways into monooxygenase and oxidase flavoenzymes, using an integrated computational and experimental approach. Enhanced-statistics molecular dynamics simulations reveal spontaneous protein-guided O 2 diffusion from the bulk solvent to preorganized protein cavities. The predicted protein-guided diffusion paths and the importance of key cavity residues for oxygen diffusion were verified by combining site-directed mutagenesis, rapid kinetics experiments, and high-resolution X-ray structures. This study indicates that monooxygenase and oxidase flavoenzymes employ multiple funnel-shaped diffusion pathways to absorb O 2 from the solvent and direct it to the reacting C4a atom of the flavin cofactor. The difference in O 2 reactivity among dehydrogenases, monooxygenases, and oxidases ultimately resides in the fine modulation of the local environment embedding the reactive locus of the flavin. |
| Databáze: | OpenAIRE |
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