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

Anisha M Perez1, Marcella M Gomez2, Prashant Kalvapalle3

  • 1Department of Biosciences, Rice University, Houston, TX, USA.

Molecular Systems Biology
|December 24, 2017
PubMed

Insights

Researchers developed a new physiological model to predict the biochemical properties of the major facilitator superfamily (MFS) tetracycline efflux pump (TetB) using cellular fitness data. This approach bypasses challenging in vitro studies for antimicrobial resistance research.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Microbiology

Background:

  • Major facilitator superfamily (MFS) efflux pumps are key players in antimicrobial resistance.
  • Studying MFS proteins in vitro is difficult due to their membrane environment.

Purpose of the Study:

  • To develop a physiological model using cellular fitness to predict MFS efflux pump biochemical properties.
  • To characterize the tetracycline efflux pump TetB and its variants.

Main Methods:

  • Utilized cellular fitness as a proxy to infer biochemical parameters.
  • Developed a model to predict K(m) and V(max) for TetB and its variants.
  • Integrated in vivo protein levels for refined parameter prediction.

Main Results:

  • Successfully determined lumped biochemical parameters (K(m), V(max)) for TetB and variants.
  • Achieved more specific predictions of substrate binding affinity and pumping efficiency.
  • Demonstrated model utility by assaying a library of tet(B) variants using fitness data.

Conclusions:

  • Cellular fitness can be effectively used as a proxy to predict MFS efflux pump biochemical properties.
  • The developed model offers a novel approach to study antimicrobial resistance mechanisms.
  • This method facilitates the biochemical characterization of efflux pumps and their variants.
Keywords:
antibiotic resistanceefflux pumpmajor facilitator superfamilystructure functiontetracycline

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