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PMID: 2164211 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

lac permease of Escherichia coli: topology and sequence elements promoting membrane insertion.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 13 ·1990-07-00 ·Pages 4937-41

Calamia J, Manoil C

Abstract

The membrane topology of Escherichia coli lac permease was analyzed using a set of 36 lac permease-alkaline phosphatase (lacY-phoA) gene fusions. The level of enzymatic activity of alkaline phosphatase fused to a cytoplasmic membrane protein appears to reflect whether the fusion junction site normally faces the cytoplasm or periplasm. The alkaline phosphatase activities of cells expressing the lacY-phoA fusions distinguish between models previously proposed for the topology of lac permease and favor one with 12 transmembrane segments. This model is fully compatible with the results of earlier biochemical and immunological studies. The properties of fusions with junctions spanning two of the transmembrane segments at 2- or 3-amino acid intervals indicate that approximately half of the residues of either segment (9-11 amino acids) suffices to promote alkaline phosphatase translocation across the membrane. The additional transmembrane segment amino acids that are not required for this membrane insertion process may normally be needed in unfused lac permease after insertion for stable association with the membrane.

MeSH Terms
Alkaline Phosphatase/genetics,isolation & purification,metabolism Amino Acid Sequence Cell Membrane/enzymology Cloning, Molecular DNA Transposable Elements Escherichia coli/enzymology,genetics Escherichia coli Proteins Gene Expression Genes, Bacterial Membrane Transport Proteins/genetics,isolation & purification,metabolism Models, Structural Molecular Sequence Data Monosaccharide Transport Proteins Plasmids Protein Conformation Recombinant Fusion Proteins/isolation & purification,metabolism Restriction Mapping Symporters
Chemicals
DNA Transposable Elements Escherichia coli Proteins LacY protein, E coli Membrane Transport Proteins Monosaccharide Transport Proteins Recombinant Fusion Proteins Symporters lactose permease Alkaline Phosphatase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Calamia J
Department of Genetics, University of Washington, Seattle 98195.
Manoil C
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24 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.87.13.4937
Published
1990-07-00
Pages
4937-41
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC54236
Subset
IM
Grants
NIGMS NIH HHS · GM36896 · United States
External Links
PubMed source DOI: 10.1073/pnas.87.13.4937 PMC Full Text View journal details
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