title: Ciprofloxacin interactions with bacterial protein OmpF: modelling of FRET from a multi-tryptophan protein trimer.
abstract: 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 (K(B)). log (K(B)) 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.
authors: Centro de Química-Física Molecular, Instituto Superior Técnico, Lisbon, Portugal.
authors: Fábio
authors: F
authors: Fernandes
authors: Patrícia
authors: P
authors: Neves
authors: Paula
authors: P
authors: Gameiro
authors: Luís M S
authors: LM
authors: Loura
authors: Manuel
authors: M
authors: Prieto
mesh_terms: D000890:Anti-Infective Agents / Q000737:chemistry*; D002939:Ciprofloxacin / Q000737:chemistry*; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D008081:Liposomes / Q000737:chemistry; D008962:Models, Theoretical; D018272:Porins / Q000737:chemistry*; D014364:Tryptophan / Q000737:chemistry*
chemical_list: D000890:Anti-Infective Agents; D008081:Liposomes; C109237:OmpF protein; D018272:Porins; D002939:Ciprofloxacin; D014364:Tryptophan; D004134:Dimyristoylphosphatidylcholine