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PMID: 9214624 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Subunit interactions in ABC transporters: a conserved sequence in hydrophobic membrane proteins of periplasmic permeases defines an important site of interaction with the ATPase subunits.

The EMBO journal ·Vol. 16 ·No. 11 ·1997-06-02 ·Pages 3066-77

Mourez M, Hofnung M, Dassa E

Abstract

The cytoplasmic membrane proteins of bacterial binding protein-dependent transporters belong to the superfamily of ABC transporters. The hydrophobic proteins display a conserved, at least 20 amino acid EAA---G---------I-LP region exposed in the cytosol, the EAA region. We mutagenized the EAA regions of MalF and MalG proteins of the Escherichia coli maltose transport system. Substitutions at the same positions in MalF and MalG have different phenotypes, indicating that EAA regions do not act symmetrically. Mutations in malG or malF that slightly affect or do not affect transport, determine a completely defective phenotype when present together. This suggests that EAA regions of MalF and MalG may interact during transport. Maltose-negative mutants fall into two categories with respect to the cellular localization of the MalK ATPase: in the first, MalK is membrane-bound, as in wild-type strains, while in the second, it is cytosolic, as in strains deleted in the malF and malG genes. From maltose-negative mutants of the two categories, we isolated suppressor mutations within malK that restore transport. They map mainly in the putative helical domain of MalK, suggesting that EAA regions may constitute a recognition site for the ABC ATPase helical domain.

MeSH Terms
ATP-Binding Cassette Transporters/genetics,metabolism Adenosine Triphosphatases/metabolism Bacterial Proteins/genetics,metabolism Biological Transport Carrier Proteins/genetics,metabolism Cell Compartmentation Cell Membrane/metabolism Conserved Sequence Cytoplasm/chemistry Escherichia coli/physiology Escherichia coli Proteins Maltose/metabolism Maltose-Binding Proteins Monosaccharide Transport Proteins Mutagenesis, Site-Directed Mutation Periplasmic Binding Proteins Phenotype Protein Binding Protein Conformation
Chemicals
ATP-Binding Cassette Transporters Bacterial Proteins Carrier Proteins Escherichia coli Proteins MalE protein, E coli MalG protein, E coli MalK protein, Bacteria MalK protein, E coli Maltose-Binding Proteins Monosaccharide Transport Proteins Periplasmic Binding Proteins maltose transport system, E coli Maltose Adenosine Triphosphatases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Mourez M
Unité de Programmation Moléculaire et Toxicologie Génétique, CNRS URA 1444, Institut Pasteur, Paris, France.
Hofnung M
Dassa E
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/16.11.3066
Published
1997-06-02
Pages
3066-77
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1169925
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/16.11.3066 PMC Full Text View journal details
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