Altered substrate selection of the melibiose transporter (MelY) of Enterobacter cloacae involving point mutations in Leu-88, Leu-91, and Ala-182 that confer enhanced maltose transport.

Journal of Bacteriology
Steven G ShinnickManuel F Varela

Abstract

We isolated mutants of Escherichia coli HS4006 containing the melibiose-H(+) symporter (MelY) from Enterobacter cloacae that had enhanced fermentation on 1% maltose MacConkey plates. DNA sequencing revealed three site classes of mutations: L-88-P, L-91-P, and A-182-P. The mutants L-88-P and L-91-P had 3.6- and 5.1-fold greater maltose uptake than the wild type and enhanced apparent affinities for maltose. Energy-coupled transport was defective for melibiose accumulation, but detectable maltose accumulation for the mutants indicated that active transport is dependent upon the substrate transported through the carrier. We conclude that the residues Leu-88, Leu-91 (transmembrane segment 3 [TMS-3]), and Ala-182 (TMS-6) of MelY mediate sugar selection. These data represent the first MelY mutations that confer changes in sugar selection.

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Citations

Mar 19, 2009·The Journal of Membrane Biology·Yang PengManuel F Varela
Jan 8, 2008·Journal of Molecular Biology·Sotiria Tavoulari, Stathis Frillingos
Apr 24, 2016·Applied and Environmental Microbiology·Ramanan SekarThomas J DiChristina
Nov 24, 2004·The Journal of Biological Chemistry·Jason A Hall, Peter C Maloney

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