Ciprofloxacin interactions with bacterial protein OmpF: Modelling of FRET from a multi-tryptophan protein trimer
| Autor: | Patrícia Neves, Manuel Prieto, Paula Gameiro, Fábio Fernandes, Luís M. S. Loura |
|---|---|
| Jazyk: | angličtina |
| Předmět: |
Biophysics
Porins Trimer Biochemistry Fluorescence 03 medical and health sciences Anti-Infective Agents Ciprofloxacin Fluorescence Resonance Energy Transfer Binding site Lipid bilayer 030304 developmental biology 0303 health sciences Quenching (fluorescence) 030306 microbiology Chemistry Bilayer Porin Tryptophan technology industry and agriculture Cell Biology Models Theoretical biochemical phenomena metabolism and nutrition Binding constant Crystallography Förster resonance energy transfer Membrane model system Membrane protein Liposomes bacteria lipids (amino acids peptides and proteins) Dimyristoylphosphatidylcholine Fluoroquinolones |
| Zdroj: | Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP |
| ISSN: | 0005-2736 |
| DOI: | 10.1016/j.bbamem.2007.07.016 |
| Popis: | The outer membrane protein F (OmpF) is known to play an important role in the uptake of fluoroquinolone antibiotics by bacteria. In this study, the degree of binding of the fluoroquinolone antibiotic ciprofloxacin to OmpF in a lipid membrane environment is quantified using a methodology based on Förster resonance energy transfer (FRET). Analysis of the fluorescence quenching of OmpF is complex as each OmpF monomer presents two tryptophans at different positions, thus sensing two different distributions of acceptors in the bilayer plane. Specific FRET formalisms were derived accounting for the different energy transfer contributions to quenching of each type of tryptophan of OmpF, allowing the recovery of upper and lower boundaries for the ciprofloxacin-OmpF binding constant (KB). log (KB) was found to lie in the range 3.15-3.62 or 3.58-4.00 depending on the location for the ciprofloxacin binding site assumed in the FRET modelling, closer to the centre or to the periphery of the OmpF trimer, respectively. This methodology is suitable for the analysis of FRET data obtained with similar protein systems and can be readily adapted to different geometries. http://www.sciencedirect.com/science/article/B6T1T-4PCPFM2-2/1/3947bd4e257b7a6021d5f6b559a91dc3 |
| Databáze: | OpenAIRE |
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