عنوان

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

Number

LA3z80f2pr

Title Proper

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

General Material Designation

[Article]

First Statement of Responsibility

Perez, Anisha M; Gomez, Marcella M; Kalvapalle, Prashant; O'Brien-Gilbert, Erin; Bennett, Matthew R; Shamoo, Yousif

Text of Note

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 Km and Vmax 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.

Date of Publication

2017

Title

UC Santa Cruz

Electronic name

[Article]

275578

a

Y