Purification and Initial Characterization of a Putative Blue Light Regulated Phosphodiesterase from Escherichia coli
| Autor: | Jason Key, Keith Moffat, Sudarshan Rajagopal, David J. Boerema, Erin B. Purcell |
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| Rok vydání: | 2004 |
| Předmět: |
Light
Molecular Sequence Data Color Gene Expression Context (language use) Biology medicine.disease_cause Biochemistry chemistry.chemical_compound Flavins EAL domain Escherichia coli medicine Amino Acid Sequence Cloning Molecular Physical and Theoretical Chemistry BLUF domain Flavin adenine dinucleotide Sequence Homology Amino Acid Phosphoric Diester Hydrolases Spectrum Analysis Phosphodiesterase General Medicine Hydrogen-Ion Concentration Chromophore biology.organism_classification chemistry Chromatography Gel Sequence Alignment Bacteria |
| Zdroj: | Photochemistry and Photobiology. |
| ISSN: | 0031-8655 |
| Popis: | The Escherichia coli protein YcgF contains a photosensory flavin adenine dinucleotide (FAD)-binding BLUF domain covalently linked to an EAL domain, which is predicted to have cyclic-di-guanosine monophosphate (GMP) phosphodiesterase activity. We have cloned, overexpressed and purified this protein, which we refer to as blue light-regulated phosphodiesterase (Blrp) for its putative activity. Blrp undergoes a reversible photocycle after exposure to light in which the spectrum of its photostationary state and kinetics of recovery of the dark state are similar to those of the isolated BLUF domain of the AppA protein. Unlike the AppA BLUF domain, the chromophore environment in the context of full-length Blrp is asymmetric, and the protein does not undergo any detectable global changes on exposure to blue light. When overexpressed in E. coli, Blrp copurifies with certain proteins which suggests that it plays a protective role in response to oxidative stress. Predicted proteins from Klebsiella pneumoniae and from a bacterium in the Sargasso Sea are similar to E. coli Blrp in both their BLUF and EAL domains, which suggests that blue light sensing in these bacteria may follow similar pathways. |
| Databáze: | OpenAIRE |
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