title: KpsC and KpsS are retaining 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) transferases involved in synthesis of bacterial capsules.
abstract: Capsular polysaccharides (CPSs) are high-molecular-mass cell-surface polysaccharides, that act as important virulence factors for many pathogenic bacteria. Several clinically important Gram-negative pathogens share similar systems for CPS biosynthesis and export; examples include Escherichia coli, Campylobacter jejuni, Haemophilus influenzae, Neisseria meningitidis, and Pasteurella multocida. Each CPS contains a serotype-specific repeat-unit structure, but the glycans all possess a lipid moiety at their reducing termini. In E. coli and N. meningitidis, the predominant lipid is a lysophosphatidylglycerol moiety that is attached to the repeat-unit domain of the CPS via multiple residues of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo), referred to as a poly-Kdo linker. The Kdo residues are β-linked, suggesting that they are synthesized by retaining glycosyltransferases. To date, the only characterized Kdo transferases are the inverting enzymes that catalyze the α-linkages found in lipopolysaccharide. Here, we identify two conserved proteins from CPS assembly systems, KpsC and KpsS, as the β-Kdo-transferases and demonstrate in vitro reconstitution of poly-Kdo linker assembly on a fluorescent phosphatidylglycerol acceptor. KpsS adds the first Kdo residue, and this reaction product is then extended by KpsC. Cross-complementation experiments demonstrate that the E. coli and N. meningitidis protein homologs are functionally conserved.
authors.affiliation: Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada N1G 2W1.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Willis
authors.forename: Chris
authors.initials: C
authors.lastname: Whitfield
mesh_terms: D016667:Bacterial Capsules / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D005816:Genetic Complementation Test; D009345:Neisseria meningitidis / Q000235:genetics / Q000378:metabolism*; D013400:Sugar Acids / Q000378:metabolism; D014166:Transferases / Q000235:genetics / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D013400:Sugar Acids; C002532:2-keto-3-deoxyoctonate; D014166:Transferases; C019917:3-deoxy-D-manno-octulosonate transferase