Understanding transport by the major facilitator superfamily (MFS): structures pave the way

Quistgaard, E.M.; Löw, Christian; Guettou, Fatma; Nordlund, P.

doi:10.1038/nrm.2015.25

Cited by 382 publications

(441 citation statements)

References 77 publications

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“…There are three groups of four alpha helices that interact to form the canonical MFS fold. Here, we use the nomenclature from Quistgaard et al , 2016 for the helices. A‐helices shown in blue (TM1, TM4, TM7, and TM10) are positioned at the center of the transporter and make up the transport path needed for substrate binding and exchange or cotransport coupling.…”

Section: Resultsmentioning

confidence: 99%

“…In addition to TetB, there are many other MFS transporters, which contribute to single and multidrug resistance in a wide variety of pathogens and their thorough characterization is of great interest (Fluman & Bibi, 2009; Alegre et al , 2016). Despite the low percentage of protein sequence similarity among MFS transporters, their overall structure is largely conserved with a general understanding of how specific regions of the transporter contribute to substrate binding and/or pumping efficiency (Yan, 2015; Quistgaard et al , 2016). …”

Section: Introductionmentioning

confidence: 99%

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Using cellular fitness to map the structure and function of a major facilitator superfamily effluxer

Perez

Gomez

Kalvapalle

et al. 2017

Molecular Systems Biology

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The major facilitator superfamily (MFS) effluxers are prominent mediators of antimicrobial resistance. The biochemical characterization of MFS proteins is hindered by their complex membrane environment that makes in vitro biochemical analysis challenging. Since the physicochemical properties of proteins drive the fitness of an organism, we posed the question of whether we could reverse that relationship and derive meaningful biochemical parameters for a single protein simply from fitness changes it confers under varying strengths of selection. Here, we present a physiological model that uses cellular fitness as a proxy to predict the biochemical properties of the MFS tetracycline efflux pump, TetB, and a family of single amino acid variants. We determined two lumped biochemical parameters roughly describing K m and V max for TetB and variants. Including in vivo protein levels into our model allowed for more specified prediction of pump parameters relating to substrate binding affinity and pumping efficiency for TetB and variants. We further demonstrated the general utility of our model by solely using fitness to assay a library of tet(B) variants and estimate their biochemical properties.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Using cellular fitness to map the structure and function of a major facilitator superfamily effluxer

Perez

Gomez

Kalvapalle

et al. 2017

Molecular Systems Biology

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show abstract

“…Such problems include the fact that it can be difficult to solubilize membrane-associated proteins for crystallization, and that X-ray crystallography relies on the arrangement of protein into a regular assembly. This often means that only single conformations can be sampled, leaving much of the dynamic cycle of these proteins unobserved and continually debated [1]. Additionally, membrane transporters may consist of large proteins, or can exist as multidomain complexes making them difficult to measure for size-limited methods such as NMR spectroscopy (though recent advances in solid state NMR are starting to minimize this limitation) [2].…”

Section: Introductionmentioning

confidence: 99%

Membrane transporters studied by EPR spectroscopy: structure determination and elucidation of functional dynamics

Mullen

Hall

Diegel

et al. 2016

Biochemical Society Transactions

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During their mechanistic cycles membrane transporters often undergo extensive conformational changes, sampling a range of orientations, in order to complete their function. Such membrane transporters present somewhat of a challenge to conventional structural studies; indeed, crystallization of membrane-associated proteins sometimes require conditions that vary vastly from their native environments. Moreover, this technique currently only allows for visualization of single selected conformations during any one experiment. EPR spectroscopy is a magnetic resonance technique that offers a unique opportunity to study structural, environmental and dynamic properties of such proteins in their native membrane environments, as well as readily sampling their substrate-binding-induced dynamic conformational changes especially through complementary computational analyses. Here we present a review of recent studies that utilize a variety of EPR techniques in order to investigate both the structure and dynamics of a range of membrane transporters and associated proteins, focusing on both primary (ABC-type transporters) and secondary active transporters which were key interest areas of the late Professor Stephen Baldwin to whom this review is dedicated.

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“…MFS membrane transport proteins are conserved within both prokaryotic and eukaryotic organisms and have high structural similarity and yet divergent functions and substrate specificities (48). MFS transporters typically move one substrate by coupling transport, with a second substrate diffusing its concentration gradient, such as a proton.…”

Section: Figmentioning

confidence: 99%

A New Natural Product Analog of Blasticidin S Reveals Cellular Uptake Facilitated by the NorA Multidrug Transporter

Davison

Lohith

Wang

et al. 2017

Antimicrob Agents Chemother

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The permeation of antibiotics through bacterial membranes to their target site is a crucial determinant of drug activity but in many cases remains poorly understood. During screening efforts to discover new broad-spectrum antibiotic compounds from marine sponge samples, we identified a new analog of the peptidyl nucleoside antibiotic blasticidin S that exhibited up to 16-fold-improved potency against a range of laboratory and clinical bacterial strains which we named P10. Whole-genome sequencing of laboratory-evolved strains of Staphylococcus aureus resistant to blasticidin S and P10, combined with genome-wide assessment of the fitness of barcoded Escherichia coli knockout strains in the presence of the antibiotics, revealed that restriction of cellular access was a key feature in the development of resistance to this class of drug. In particular, the gene encoding the well-characterized multidrug efflux pump NorA was found to be mutated in 69% of all S. aureus isolates resistant to blasticidin S or P10. Unexpectedly, resistance was associated with inactivation of norA, suggesting that the NorA transporter facilitates cellular entry of peptidyl nucleosides in addition to its known role in the efflux of diverse compounds, including fluoroquinolone antibiotics.

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Understanding transport by the major facilitator superfamily (MFS): structures pave the way

Cited by 382 publications

References 77 publications

Using cellular fitness to map the structure and function of a major facilitator superfamily effluxer

Using cellular fitness to map the structure and function of a major facilitator superfamily effluxer

Membrane transporters studied by EPR spectroscopy: structure determination and elucidation of functional dynamics

A New Natural Product Analog of Blasticidin S Reveals Cellular Uptake Facilitated by the NorA Multidrug Transporter

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