The sodium-dependent D-glucose transport protein of Helicobacter pylori.
Helicobacter pylori is a gram-negative pathogenic microaerophile with a particular tropism for the mucosal surface of the gastric epithelium. Despite its obligatory microaerophilic character, it can metabolize D-glucose and/or D-galactose in both oxidative and fermentative pathways via a Na(+)-dependent secondary active transport, a glucokinase and enzymes of the pentose phosphate pathway. We have assigned the Na(+)-dependent transport of glucose to the protein product of the H. pylori 1174 gene. The gene was heterologously expressed in a glucose transport-deficient Escherichia coli strain, where transport activities of radiolabelled D-glucose, D-galactose and 2-deoxy-D-glucose were restored, consistent with the expected specificity of the hexose uptake system in H. pylori. D-mannose was also identified as a substrate. The HP1174 transport protein was purified and reconstituted into proteoliposomes, where sodium dependence of sugar transport activity was demonstrated. Additionally the tryptophan/tyrosine fluorescence of the purified protein showed quenching by 2-deoxy-D-glucose, D-mannose, D-glucose or D-galactose in the presence of sodium ions. This is the first reported purification and characterization of an active glucose transport protein member of the TC 2.1.7 subgroup of the Major Facilitator Superfamily, constituting the route for entry of sugar nutrients into H. pylori. A model is derived of its three-dimensional structure as a paradigm of the family.
Psakis
Georgios
G
The Astbury Centre for Structural Molecular Biology, Institute of Membrane and Systems Biology, University of Leeds, Leeds LS2 9JT, UK.
Saidijam
Massoud
M
Shibayama
Keigo
K
Polaczek
Julia
J
Bettaney
Kim E
KE
Baldwin
Jocelyn M
JM
Baldwin
Stephen A
SA
Hope
Ryan
R
Essen
Lars-Oliver
LO
Essenberg
Richard C
RC
Henderson
Peter J F
PJ
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001693:Biological Transport, Active; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D005947:Glucose / Q000378:metabolism; D016480:Helicobacter pylori / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D016296:Mutagenesis; D017433:Protein Structure, Secondary; D012964:Sodium / Q000378:metabolism; D051247:Sodium-Glucose Transport Proteins / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D051247:Sodium-Glucose Transport Proteins; D012964:Sodium; D005947:Glucose