Sugar recognition by the lactose permease of Escherichia coli

José Luis Vázquez-Ibari, Lan Guan, Adam B. Weinglass, Gill Verner, Ruth Gordillo, H. Ronald Kaback

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Biochemical, luminescence and mass spectroscopy approaches indicate that Trp-151 (helix V) plays an important role in hydrophobic stacking with the galactopyranosyl ring of substrate and that Glu-269 (helix VIII) is essential for substrate affinity and specificity. The x-ray structure of the lactose permease (LacY) with bound substrate is consistent with these conclusions and suggests that a possible H-bond between Glu-269 and Trp-151 may play a critical role in the architecture of the binding site. We have now probed this relationship by exploiting the intrinsic luminescence of a single Trp-151 LacY with various replacements for Glu-269. Mutations at position 269 dramatically alter the environment of Trp-151 in a manner that correlates with binding affinity of LacY substrates. Furthermore, chemical modification of Trp-151 with N-bromosuccinimide indicates that Glu-269 forms an H-bond with the indole N. It is concluded that 1) an H-bond between the indole N and Glu-269 optimizes the formation of the substrate binding site in the inward facing conformation of LacY, and 2) the disposition of the residues implicated in sugar binding in different conformers suggests that sugar binding by LacY involves induced fit.

Original languageEnglish (US)
Pages (from-to)49214-49221
Number of pages8
JournalJournal of Biological Chemistry
Volume279
Issue number47
DOIs
StatePublished - Nov 19 2004

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Luminescence
Sugars
Escherichia coli
Binding Sites
Bromosuccinimide
Substrates
Substrate Specificity
Mass Spectrometry
X-Rays
Mutation
Chemical modification
Conformations
lactose permease
indole
Spectroscopy
X rays

ASJC Scopus subject areas

  • Biochemistry

Cite this

Vázquez-Ibari, J. L., Guan, L., Weinglass, A. B., Verner, G., Gordillo, R., & Kaback, H. R. (2004). Sugar recognition by the lactose permease of Escherichia coli. Journal of Biological Chemistry, 279(47), 49214-49221. https://doi.org/10.1074/jbc.M407408200

Sugar recognition by the lactose permease of Escherichia coli. / Vázquez-Ibari, José Luis; Guan, Lan; Weinglass, Adam B.; Verner, Gill; Gordillo, Ruth; Kaback, H. Ronald.

In: Journal of Biological Chemistry, Vol. 279, No. 47, 19.11.2004, p. 49214-49221.

Research output: Contribution to journalArticle

Vázquez-Ibari, JL, Guan, L, Weinglass, AB, Verner, G, Gordillo, R & Kaback, HR 2004, 'Sugar recognition by the lactose permease of Escherichia coli', Journal of Biological Chemistry, vol. 279, no. 47, pp. 49214-49221. https://doi.org/10.1074/jbc.M407408200
Vázquez-Ibari JL, Guan L, Weinglass AB, Verner G, Gordillo R, Kaback HR. Sugar recognition by the lactose permease of Escherichia coli. Journal of Biological Chemistry. 2004 Nov 19;279(47):49214-49221. https://doi.org/10.1074/jbc.M407408200
Vázquez-Ibari, José Luis ; Guan, Lan ; Weinglass, Adam B. ; Verner, Gill ; Gordillo, Ruth ; Kaback, H. Ronald. / Sugar recognition by the lactose permease of Escherichia coli. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 47. pp. 49214-49221.
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