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PMID: 16498400 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Two distinct but interchangeable mechanisms for flipping of lipid-linked oligosaccharides.

The EMBO journal ·Vol. 25 ·No. 5 ·2006-03-08 ·Pages 967-76

Alaimo C, Catrein I, Morf L, Marolda CL, Callewaert N, Valvano MA, Feldman MF, Aebi M

Abstract

Translocation of lipid-linked oligosaccharide (LLO) intermediates across membranes is an essential but poorly understood process in eukaryotic and bacterial glycosylation pathways. Membrane proteins defined as translocases or flippases are implicated to mediate the translocation reaction. The membrane protein Wzx has been proposed to mediate the translocation across the plasma membrane of lipopolysaccharide (LPS) O antigen subunits, which are assembled on an undecaprenyl pyrophosphate lipid carrier. Similarly, PglK (formerly WlaB) is a Campylobacter jejuni-encoded ABC-type transporter proposed to mediate the translocation of the undecaprenylpyrophosphate-linked heptasaccharide intermediate involved in the recently identified bacterial N-linked protein glycosylation pathway. A combination of genetic and carbohydrate structural analyses defined and characterized flippase activities in the C. jejuni N-linked protein glycosylation and the Escherichia coli LPS O antigen biosynthesis. PglK displayed relaxed substrate specificity with respect to the oligosaccharide structure of the LLO intermediate and complemented a wzx deficiency in E. coli O-antigen biosynthesis. Our experiments provide strong genetic evidence that LLO translocation across membranes can be catalyzed by two distinct proteins that do not share any sequence similarity.

MeSH Terms
ATP-Binding Cassette Transporters/genetics,metabolism Bacterial Proteins/genetics,metabolism Campylobacter jejuni/chemistry,genetics,metabolism DNA, Recombinant/genetics Escherichia coli/chemistry,genetics,metabolism Escherichia coli Proteins/genetics,metabolism Gene Expression Regulation, Bacterial Genetic Complementation Test Glycosylation Lipopolysaccharides/chemistry,metabolism Membrane Transport Proteins/genetics,metabolism Mutation/genetics O Antigens/metabolism
Chemicals
ATP-Binding Cassette Transporters Bacterial Proteins DNA, Recombinant Escherichia coli Proteins Lipopolysaccharides Membrane Transport Proteins O Antigens WzxE protein, E coli lipid-linked oligosaccharides wlaB protein, Campylobacter jejuni
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Alaimo Cristina
Department of Biology, Institute of Microbiology, Swiss Federal Institute of Technology, ETH-Hönggerberg, Zurich, Switzerland.
Catrein Ina
Morf Laura
Marolda Cristina L
Callewaert Nico
Valvano Miguel A
Feldman Mario F
Aebi Markus
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1038/sj.emboj.7601024
Published
2006-03-08
Epub
2006-00-23
Pages
967-76
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1409731
Subset
IM
External Links
PubMed source DOI: 10.1038/sj.emboj.7601024 PMC Full Text View journal details
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