Figure 4

Variants of the E. coli GabP that exhibit Δ(TSR) phenotypes Using data analogous to Figure 1, the concentrations of competing substrates were adjusted empirically such that the initial rates of label accumulation were superimposed for E. coli strains expressing the "control" gab permease (GabP). As a result, any separation between initial rate uptake curves for [¹⁴C]GABA (▲) and [³H]NA (■) provides a highly intuitive visual representation of a Δ(TSR) phenotype. Panel N302C shows TSR analysis of the single-Cys GabP variant, N302C. Compared to the Cys-less GabP control (TSR = 8) for which the initial label accumulation rates are superimposed [5], the N302C shows a relative increase in the specificity for NA with a calculated TSR of 2.5. The Panel INS Ala 320 shows TSR analysis of the GabP variant, INS Ala 320, which has an extra alanine residue inserted at position 320. Compared to the wild type GabP control (TSR = 4) for which the initial label accumulation rates are superimposed [6], the INS Ala 320 exhibits a relative increase in specificity for GABA (i.e., opposite of the Panel N302C) with a calculated TSR of 16.