A Structural Basis for the Regulation of an H-NOX-Associated Cyclic-di-GMP Synthase/Phosphodiesterase Enzyme by Nitric Oxide-Bound H-NOX
| Autor: | Daniel P. Raleigh, Elizabeth M. Boon, Bowu Luan, Tanaya Lahiri |
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| Rok vydání: | 2014 |
| Předmět: |
Models
Molecular inorganic chemicals Cyclic di-GMP Shewanella Biochemistry & Molecular Biology Heme Medical Biochemistry and Metabolomics Nitric Oxide Biochemistry Article Nitric oxide Medicinal and Biomolecular Chemistry 03 medical and health sciences chemistry.chemical_compound Bacterial Proteins Models Site-Directed Cyclic GMP 030304 developmental biology 0303 health sciences Molecular Structure biology ATP synthase Phosphoric Diester Hydrolases 030306 microbiology Escherichia coli Proteins Biofilm Molecular Phosphodiesterase Enzyme assay Oxygen chemistry Mutagenesis Biofilms Mutagenesis Site-Directed biology.protein Mutant Proteins Diguanylate cyclase Biochemistry and Cell Biology Phosphorus-Oxygen Lyases Oxygen binding |
| Zdroj: | Biochemistry, vol 53, iss 13 Biochemistry |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi401597m |
| Popis: | Biofilms are surface-attached communities of bacteria enclosed in a polysaccharide matrix. Bacteria in a biofilm are extremely resistant to antibiotics. Several recent reports have linked the signaling molecule nitric oxide (NO) with biofilm dispersal. We have previously reported that an H-NOX (heme-nitric oxide/oxygen binding) protein in the biofilm-dwelling bacterium Shewanella woodyi mediates NO-induced biofilm dispersal. In S. woodyi, H-NOX (SwH-NOX) is cocistronic with a gene encoding a dual-functioning diguanylate cyclase/phosphodiesterase enzyme, designated here as HaCE (H-NOX-associated cyclic-di-GMP processing enzyme). Enzymes such as these are responsible for regulating the intracellular concentrations of cyclic-di-GMP, a secondary signaling molecule essential to biofilm formation in bacteria. We have demonstrated that NO-bound SwH-NOX regulates both enzymatic activities of SwHaCE, resulting in decreased cellular cyclic-di-GMP levels and disruption of biofilm formation. Thus, H-NOX/HaCE represents a potential drug target for regulating biofilm formation. In this work, the SwH-NOX surface residues critical for the formation of a protein complex with SwHaCE are identified using nuclear magnetic resonance, fluorescence quenching, and cosedimentation. Enzyme assays confirm this protein–protein interface and its importance for H-NOX/HaCE function. |
| Databáze: | OpenAIRE |
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