Abstract
The overexpression of multidrug efflux pumps is an important mechanism of clinical resistance in Gram-negative bacteria. Recently, four small molecules were discovered that inhibit efflux in Escherichia coli and interact with the AcrAB-TolC efflux pump component AcrA. However, the binding site(s) for these molecules was not determined. Here, we combine ensemble docking and molecular dynamics simulations with tryptophan fluorescence spectroscopy, site-directed mutagenesis, and antibiotic susceptibility assays to probe binding sites and effects of binding of these molecules. We conclude that clorobiocin and SLU-258 likely bind at a site located between the lipoyl and β-barrel domains of AcrA.
| Original language | English |
|---|---|
| Pages (from-to) | 648-658 |
| Number of pages | 11 |
| Journal | Biophysical Journal |
| Volume | 116 |
| Issue number | 4 |
| DOIs | |
| State | Published - 19 Feb 2019 |
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Identification of Binding Sites for Efflux Pump Inhibitors of the AcrAB-TolC Component AcrA. / Darzynkiewicz, Zbigniew M.; Green, Adam T.; Abdali, Narges; Hazel, Anthony; Fulton, Ronnie L.; Kimball, Joseph; Gryczynski, Zygmunt; Gumbart, James C.; Parks, Jerry M.; Smith, Jeremy C.; Zgurskaya, Helen I.
In: Biophysical Journal, Vol. 116, No. 4, 19.02.2019, p. 648-658.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Identification of Binding Sites for Efflux Pump Inhibitors of the AcrAB-TolC Component AcrA
AU - Darzynkiewicz, Zbigniew M.
AU - Green, Adam T.
AU - Abdali, Narges
AU - Hazel, Anthony
AU - Fulton, Ronnie L.
AU - Kimball, Joseph
AU - Gryczynski, Zygmunt
AU - Gumbart, James C.
AU - Parks, Jerry M.
AU - Smith, Jeremy C.
AU - Zgurskaya, Helen I.
PY - 2019/2/19
Y1 - 2019/2/19
N2 - The overexpression of multidrug efflux pumps is an important mechanism of clinical resistance in Gram-negative bacteria. Recently, four small molecules were discovered that inhibit efflux in Escherichia coli and interact with the AcrAB-TolC efflux pump component AcrA. However, the binding site(s) for these molecules was not determined. Here, we combine ensemble docking and molecular dynamics simulations with tryptophan fluorescence spectroscopy, site-directed mutagenesis, and antibiotic susceptibility assays to probe binding sites and effects of binding of these molecules. We conclude that clorobiocin and SLU-258 likely bind at a site located between the lipoyl and β-barrel domains of AcrA.
AB - The overexpression of multidrug efflux pumps is an important mechanism of clinical resistance in Gram-negative bacteria. Recently, four small molecules were discovered that inhibit efflux in Escherichia coli and interact with the AcrAB-TolC efflux pump component AcrA. However, the binding site(s) for these molecules was not determined. Here, we combine ensemble docking and molecular dynamics simulations with tryptophan fluorescence spectroscopy, site-directed mutagenesis, and antibiotic susceptibility assays to probe binding sites and effects of binding of these molecules. We conclude that clorobiocin and SLU-258 likely bind at a site located between the lipoyl and β-barrel domains of AcrA.
UR - http://www.scopus.com/inward/record.url?scp=85060472928&partnerID=8YFLogxK
U2 - 10.1016/j.bpj.2019.01.010
DO - 10.1016/j.bpj.2019.01.010
M3 - Article
VL - 116
SP - 648
EP - 658
JO - Biophysical Journal
JF - Biophysical Journal
SN - 0006-3495
IS - 4
ER -