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301.
Zbigniew M. Darzynkiewicz Adam T. Green Narges Abdali Anthony Hazel Ronnie L. Fulton Joseph Kimball Zygmunt Gryczynski James C. Gumbart Jerry M. Parks Jeremy C. Smith Helen I. Zgurskaya 《Biophysical journal》2019,116(4):648-658
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. 相似文献
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Abstract Correlated measurements of total cellular RNA and DNA of cultured human keratinocytes by flow cytometry, followed by multivariate analysis, discriminate three distinct subpopulations of cells differing in RNA content. The first subpopulation is comprised of small cells resembling basal cells of epidermis, with low RNA content and long (100–300 h) generation times. The second subpopulation consists of keratinocytes resembling cells in the spinous layer of epidermis, characterized by increased RNA content and shorter (35–40 h) generation times. The third subpopulation consists of the largest, keratinohyalin-containing cells which remain in G1 and undergo terminal differentiation. In contrast to total cellular RNA, correlated measurements of DNA and nuclear RNA reveal that: (1) entrance of all cultured cells from G1 into S phase occurs only after accumulation of the same, threshold amount of nuclear RNA; hence there is only a single population of S + G2 + M-phase cells; (2) there are two distinct subpopulations in G1 , one with minimal nuclear RNA content and another with increased RNA. Stathmokinetic experiments indicate that the G1 -phase cells with low nuclear RNA have distinctly longer residence times in G1 compared to cells with high nuclear RNA content. Thus, measurements of the total cellular RNA versus nuclear RNA content reveal kinetically distinct cell subpopulations. Whereas total cellular RNA content correlates more with differentiation, nuclear RNA content reflects primarily the kinetic properties of the cell. 相似文献