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PMID: 2553664 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Export of FepA::PhoA fusion proteins to the outer membrane of Escherichia coli K-12.

Journal of bacteriology ·Vol. 171 ·No. 11 ·1989-11-00 ·Pages 5894-900

Murphy CK, Klebba PE

Abstract

A library of fepA::phoA gene fusions was generated in order to study the structure and secretion of the Escherichia coli K-12 ferric enterobactin receptor, FepA. All of the fusion proteins contained various lengths of the amino-terminal portion of FepA fused in frame to the catalytic portion of bacterial alkaline phosphatase. Localization of FepA::PhoA fusion proteins in the cell envelope was dependent on the number of residues of mature FepA present at the amino terminus. Hybrids containing up to one-third of the amino-terminal portion of FepA fractionated with their periplasm, while those containing longer sequences of mature FepA were exported to the outer membrane. Outer membrane-localized fusion proteins expressed FepA sequences on the external face of the outer membrane and alkaline phosphatase moieties in the periplasmic space. From sequence determinations of the fepA::phoA fusion joints, residues within FepA which may be exposed on the periplasmic side of the outer membrane were identified.

MeSH Terms
Alkaline Phosphatase/genetics,metabolism Bacterial Outer Membrane Proteins/genetics,metabolism Blotting, Western Carrier Proteins/genetics,metabolism Cell Membrane/metabolism Cloning, Molecular DNA Transposable Elements Escherichia coli/genetics,metabolism Genes, Bacterial Receptors, Cell Surface Recombinant Fusion Proteins/metabolism Trypsin
Chemicals
Bacterial Outer Membrane Proteins Carrier Proteins DNA Transposable Elements Receptors, Cell Surface Recombinant Fusion Proteins enterobactin receptor Alkaline Phosphatase Trypsin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Murphy C K
Department of Microbiology, Medical College of Wisconsin, Milwaukee 53226.
Klebba P E
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.171.11.5894-5900.1989
Published
1989-11-00
Pages
5894-900
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC210451
Subset
IM
Grants
NIAID NIH HHS · AI22608 · United States
External Links
PubMed source DOI: 10.1128/jb.171.11.5894-5900.1989 PMC Full Text View journal details
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