Fluorescence Quenching as a Tool to Investigate Quinolone Antibiotic Interactions with Bacterial Protein OmpF
authors Neves, P; Sousa, I; Winterhalter, M; Gameiro, P
journal JOURNAL OF MEMBRANE BIOLOGY
author keywords Spectrofluorescence; Biophysics; Structure of membrane protein; Fluorescent probe
keywords ESCHERICHIA-COLI; STAPHYLOCOCCUS-AUREUS; PSEUDOMONAS-AERUGINOSA; PORIN; ACCUMULATION; FLUOROQUINOLONES; CIPROFLOXACIN; MOXIFLOXACIN; PERMEATION; LIPOSOMES
abstract The outer membrane porin OmpF is an important protein for the uptake of antibiotics through the outer membrane of gram-negative bacteria; however, the possible binding sites involved in this uptake are still not recognized. Determination, at the molecular level, of the possible sites of antibiotic interaction is very important, not only to understand their mechanism of action but also to unravel bacterial resistance. Due to the intrinsic OmpF fluorescence, attributed mainly to its tryptophans (Trp(214), Trp(61)), quenching experiments were used to assess the site(s) of interaction of some quinolone antibiotics. OmpF was reconstituted in different organized structures, and the fluorescence quenching results, in the presence of two quenching agents, acrylamide and iodide, certified that acrylamide quenches Trp(61) and iodide Trp(214). Similar data, obtained in presence of the quinolones, revealed distinct behaviors for these antibiotics, with nalidixic acid interacting near Trp(214) and moxifloxacin near Trp(61). These studies, based on straightforward and quick procedures, show the existence of conformational changes in the protein in order to adapt to the different organized structures and to interact with the quinolones. The extent of reorganization of the protein in the presence of the different quinolones allowed an estimate on the sites of protein/quinolone interaction.
publisher SPRINGER
issn 0022-2631
year published 2009
volume 227
issue 3
beginning page 133
ending page 140
digital object identifier (doi) 10.1007/s00232-008-9152-2
web of science category Biochemistry & Molecular Biology; Cell Biology; Physiology
subject category Biochemistry & Molecular Biology; Cell Biology; Physiology
unique article identifier WOS:000263916400004
  ciceco authors
  impact metrics
journal analysis (jcr 2017):
journal impact factor 1.638
5 year journal impact factor 1.689
category normalized journal impact factor percentile 19.092
dimensions (citation analysis):
altmetrics (social interaction):



 


Apoio

1suponsers_list_ciceco.jpg