PMID:16498400

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Citation

Alaimo, C, Catrein, I, Morf, L, Marolda, CL, Callewaert, N, Valvano, MA, Feldman, MF and Aebi, M (2006) Two distinct but interchangeable mechanisms for flipping of lipid-linked oligosaccharides. EMBO J. 25:967-76

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.

Links

PubMed PMC1409731 Online version:10.1038/sj.emboj.7601024

Keywords

ATP-Binding Cassette Transporters/genetics; ATP-Binding Cassette Transporters/metabolism; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Campylobacter jejuni/chemistry; Campylobacter jejuni/genetics; Campylobacter jejuni/metabolism; DNA, Recombinant/genetics; Escherichia coli/chemistry; Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Gene Expression Regulation, Bacterial; Genetic Complementation Test; Glycosylation; Lipopolysaccharides/chemistry; Lipopolysaccharides/metabolism; Membrane Transport Proteins/genetics; Membrane Transport Proteins/metabolism; Mutation/genetics; O Antigens/metabolism

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