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Structure type: oligomer
Compound class: glycolipid
Contained glycoepitopes: IEDB_141807,IEDB_151531

• Article ID: 2677
  Bhat UR, Forsberg LS, Carlson RW "Structure of lipid A component of Rhizobium leguminosarum bv. phaseoli lipopolysaccharide. Unique nonphosphorylated lipid A containing 2-amino-2-deoxygluconate, galacturonate, and glucosamine" - Journal of Biological Chemistry 269 (1994) 14402-14410
  

  The structure of lipid A from the lipopolysaccharide of Rhizobium leguminosarum bv. phaseoli (wild type strain CE3) was investigated by alkylation analysis, nuclear magnetic resonance spectroscopy, and electrospray and fast atom bombardment mass spectrometry of the de-O-acylated lipid A. The lipid A carbohydrate backbone was shown to be a trisaccharide containing galacturonic acid, glucosamine, and the unique sugar 2-amino-2-deoxygluconic acid, previously unreported in lipopolysaccharides. Nuclear magnetic resonance spectroscopy and ethylation analyses revealed that the galacturonic acid is α-1,4-linked to the glucosamine, while the amino aldonic acid residue, which may exist as the 1,5-lactone, is attached as an aglycone to the glucosamine and, thus, occupies the reducing end of the molecule. The resulting backbone is hydrophilic and analogous to the commonly observed bisphosphorylated glucosamine disaccharide from enteric bacterial lipopolysaccharides in that both the nonreducing and reducing ends carry negatively charged substituents. The fatty acids of the R. leguminosarum lipid A are attached both as O- and N-acyl substituents to glucosamine and 2-aminogluconate. All fatty acids are hydroxylated consisting of 3-hydroxymyristate (3-OH-C14.0), 3-hydroxypentadecanoate (3-OH-C15.0), 3-hydroxypalmitate (3-OH-C16.0), 3-hydroxystearate (3-OH-C18.0), and 27-hydroxyoctacosanoate (27-OH-C28.0) in the approximate mole ratio 3:0.2:1:0.6:1. Unlike lipid As from enteric bacteria, the R. leguminosarum lipid A lacks 3-acyloxyacyl substituents; however, the long chain 27-hydroxy fatty acid carries ester-linked β-hydroxybutyrate at the 27-hydroxy position. Fast atom bombardment mass spectrometry of the de-O-acylated lipid A demonstrated the presence of 2 molecular species that differ by 28 mass units due to fatty acid heterogeneity at the two amide linkages. One species carries amide-linked 3-OH-C14.0 and 3-OH-C16.0; the second species carries 3-OH-C14.0 and 3-OH-C18.0. Each molecular species also exists as the aldonolactone, yielding molecular ions at ((M+H)+)-18. The heterogeneity in the amide-linked fatty acids further distinguishes the Rhizobium lipid A from enteric lipid As.

  NCBI PubMed ID: 8182046
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Institutions: Complex Carbohydrate Research Center, University of Georgia, Athens 30605
  
   
  
  Rhizobium leguminosarum bv. phaseoli
  CSDB ID 137109 (all data & tools)

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Structure type: oligomer
Compound class: lipid A
Contained glycoepitopes: IEDB_141181,IEDB_141807,IEDB_151531,IEDB_534865

• Article ID: 3821
  Banoub JH, El Aneed A, Cohen AM, Joly N "Structural investigation of bacterial lipopolysaccharides by mass spectrometry and tandem mass spectrometry" - Mass Spectrometry Reviews 29(4) (2010) 606-650
  

  Mass spectrometric studies are now playing a leading role in the elucidation of lipopolysaccharide (LPS) structures through the characterization of antigenic polysaccharides, core oligosaccharides and lipid A components including LPS genetic modifications. The conventional MS and MS/MS analyses together with CID fragmentation provide additional structural information complementary to the previous analytical experiments, and thus contribute to an integrated strategy for the simultaneous characterization and correct sequencing of the carbohydrate moiety.

  LPS, O-antigen, lipid A, core oligosaccharide, MS and MS/MS analyses

  NCBI PubMed ID: 20589944
  Publication DOI: 10.1002/mas.20258
  Journal NLM ID: 8219702
  Publisher: Wiley
  Correspondence: joe.banoub@dfo-mpo.gc.ca
  Institutions: Fisheries and Oceans Canada, Science Branch, Special Projects, P.O. Box 5667, St. John's, Newfoundland, Canada A1C 5X1, Department of Chemistry, Memorial University of Newfoundland, St. John's, Newfoundland, Canada A1B 3V6, College of Pharmacy and Nutrition, University of Saskatchewan, Thorvaldson Building, 110 Science Place, Saskatoon, Saskatchewan, Canada S7N 5C9, Unité de Catalyse et de Chimie du Solide, Site de l'Artois—UMR CNRS 8181, I.U.T. de Béthune, Département Chimie, 1230 rue de l'Université, BP819, 62408 Béthune Cedex, France, Institute for Marine Biosciences Room 219A, (NRC-IMB), National Research Council of Canada, Government of Canada, 1411 Oxford Street, Halifax, NS, Canada B3H 3Z1
  Methods: MS/MS, MS
  
   
  
  Rhizobium etli
  CSDB ID 31124 (all data & tools)

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Structure type: oligomer
Trivial name: core oligosaccharide
Compound class: LPS
Contained glycoepitopes: IEDB_130650,IEDB_130659,IEDB_130701,IEDB_136906,IEDB_137472,IEDB_141794,IEDB_141807,IEDB_144983,IEDB_151528,IEDB_151531,IEDB_152206,IEDB_190606,IEDB_983930,SB_44,SB_67,SB_7,SB_72

• Article ID: 4076
  Muszynski A, Laus M, Kijne JW, Carlson RW "Structures of the lipopolysaccharides from Rhizobium leguminosarum RBL5523 and its UDP-glucose dehydrogenase mutant (exo5)" - Glycobiology 21(1) (2011) 55-68
  

  Rhizobial lipopolysaccharide (LPS) is required to establish an effective symbiosis with its host plant. An exo5 mutant of R. leguminosarum RBL5523, strain RBL5808, is defective in UDP-glucose (Glc) dehydrogenase that converts UDP-Glc to UDP-glucuronic acid (GlcA). This mutant is unable to synthesize either UDP-GlcA or UDP-galacturonic acid (GalA) and is unable to synthesize extracellular and capsular polysaccharides, lacks GalA in its LPS, and is defective in symbiosis (Laus et. al. 2004. J. Bacteriol.186:6617-6625). Here, we determined and compared the structures of the RBL5523 parent and RBL5808 mutant LPSs. The parent LPS core oligosaccharide (OS), as with other R. leguminosarum and R. etli strains, is a Gal(1)Man(1)GalA(3)Kdo(3) octasaccharide in which each of the GalA residues is terminally linked. The core OS from the mutant lacks all three GalA residues. Also, the parent lipid A consists of a fatty acylated GlcNGlcNonate or GlcNGlcN disaccharide that has a GalA residue at the 4'-position, typical of other R. leguminosarum and R. etli lipids A. The mutant lipid A lacks the 4' GalA residue and the proximal glycosyl residue was only present as GlcNonate. In spite of these alterations to the lipid A and core OS, the mutant was still able to synthesize an LPS containing a normal O-chain polysaccharide (OPS), but at reduced levels. The structure of the OPS of the mutant LPS was identical to that of the parent and consists of an O-acetylated →4)-α-D-Glcp-(1→3)-α-D-QuipNAc-(1→ repeating unit.

  Lipopolysaccharide, structure, symbiosis, Rhizobium leguminosarum

  NCBI PubMed ID: 20817634
  Publication DOI: 10.1093/glycob/cwq131
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: rcarlson@ccrc.uga.edu
  Institutions: Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA
  Methods: 13C NMR, 1H NMR, methylation, NMR-2D, GC-MS, HPAEC, DOC-PAGE, MALDI-TOF MS, de-O-acetylation, composition analysis, NMR-1D
  
   
  
  Rhizobium leguminosarum RBL5808
  CSDB ID 26893 (all data & tools)

Show legend Show as text

Structure type: oligomer
Trivial name: core oligosaccharide
Compound class: LPS
Contained glycoepitopes: IEDB_130650,IEDB_130659,IEDB_130701,IEDB_136906,IEDB_137472,IEDB_141794,IEDB_141807,IEDB_144983,IEDB_151528,IEDB_151531,IEDB_152206,IEDB_190606,IEDB_983930,SB_44,SB_67,SB_7,SB_72

• Article ID: 4076
  Muszynski A, Laus M, Kijne JW, Carlson RW "Structures of the lipopolysaccharides from Rhizobium leguminosarum RBL5523 and its UDP-glucose dehydrogenase mutant (exo5)" - Glycobiology 21(1) (2011) 55-68
  

  Rhizobial lipopolysaccharide (LPS) is required to establish an effective symbiosis with its host plant. An exo5 mutant of R. leguminosarum RBL5523, strain RBL5808, is defective in UDP-glucose (Glc) dehydrogenase that converts UDP-Glc to UDP-glucuronic acid (GlcA). This mutant is unable to synthesize either UDP-GlcA or UDP-galacturonic acid (GalA) and is unable to synthesize extracellular and capsular polysaccharides, lacks GalA in its LPS, and is defective in symbiosis (Laus et. al. 2004. J. Bacteriol.186:6617-6625). Here, we determined and compared the structures of the RBL5523 parent and RBL5808 mutant LPSs. The parent LPS core oligosaccharide (OS), as with other R. leguminosarum and R. etli strains, is a Gal(1)Man(1)GalA(3)Kdo(3) octasaccharide in which each of the GalA residues is terminally linked. The core OS from the mutant lacks all three GalA residues. Also, the parent lipid A consists of a fatty acylated GlcNGlcNonate or GlcNGlcN disaccharide that has a GalA residue at the 4'-position, typical of other R. leguminosarum and R. etli lipids A. The mutant lipid A lacks the 4' GalA residue and the proximal glycosyl residue was only present as GlcNonate. In spite of these alterations to the lipid A and core OS, the mutant was still able to synthesize an LPS containing a normal O-chain polysaccharide (OPS), but at reduced levels. The structure of the OPS of the mutant LPS was identical to that of the parent and consists of an O-acetylated →4)-α-D-Glcp-(1→3)-α-D-QuipNAc-(1→ repeating unit.

  Lipopolysaccharide, structure, symbiosis, Rhizobium leguminosarum

  NCBI PubMed ID: 20817634
  Publication DOI: 10.1093/glycob/cwq131
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: rcarlson@ccrc.uga.edu
  Institutions: Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA
  Methods: 13C NMR, 1H NMR, methylation, NMR-2D, GC-MS, HPAEC, DOC-PAGE, MALDI-TOF MS, de-O-acetylation, composition analysis, NMR-1D
  
   
  
  Rhizobium etli CE3, Rhizobium leguminosarum Rlv 3841, Rhizobium leguminosarum RBL5523
  CSDB ID 26894 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: lipid A, glycolipid
Contained glycoepitopes: IEDB_141807,IEDB_151531,IEDB_176772

• Article ID: 5615
  Que-Gewirth NL, Lin S, Cotter RJ, Raetz CR "An outer membrane enzyme that generates the 2-amino-2-deoxy-gluconate moiety of Rhizobium leguminosarum lipid A" - Journal of Biological Chemistry 278(14) (2003) 12109-12119
  

  The structures of Rhizobium leguminosarum and Rhizobium etli lipid A are distinct from those found in other Gram-negative bacteria. Whereas the more typical Escherichia coli lipid A is a hexa-acylated disaccharide of glucosamine that is phosphorylated at positions 1 and 4', R. etli and R. leguminosarum lipid A consists of a mixture of structurally related species (designated A-E) that lack phosphate. A conserved distal unit, comprised of a diacylated glucosamine moiety with galacturonic acid residue at position 4' and a secondary 27-hydroxyoctacosanoyl (27-OH-C28) as part of a 2' acyloxyacyl moiety, is present in all five components. The proximal end is heterogeneous, differing in the number and lengths of acyl chains and in the identity of the sugar itself. A proximal glucosamine unit is present in B and C, but an unusual 2-amino-2-deoxy-gluconate moiety is found in D-1 and E. We now demonstrate that membranes of R. leguminosarum and R. etli can convert B to D-1 in a reaction that requires added detergent and is inhibited by EDTA. Membranes of Sinorhizobium meliloti and E. coli lack this activity. Mass spectrometry demonstrates that B is oxidized in vitro to a substance that is 16 atomic mass units larger, consistent with the formation of D-1. The oxidation of the lipid A proximal unit is also demonstrated by matrix-assisted laser desorption ionization time-of-flight mass spectrometry in the positive and negative modes using the model substrate, 1-dephospho-lipid IV(A). With this material, an additional intermediate (or by product) is detected that is tentatively identified as a lactone derivative of 1-dephospho-lipid IV(A). The enzyme, presumed to be an oxidase, is located exclusively in the outer membrane of R. leguminosarum as judged by sucrose gradient analysis. To our knowledge, an oxidase associated with the outer membranes of Gram-negative bacteria has not been reported previously

  biosynthesis, lipid A, Rhizobium leguminosarum, glycolipid

  NCBI PubMed ID: 12531907
  Publication DOI: 10.1074/jbc.M300378200
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: raetz@biochem.duke.edu
  Institutions: Department of Biochemistry, Duke University Medical Center, Durham, USA, Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, USA
  Methods: 1H NMR, TLC, MALDI-TOF MS, biological assays, radiolabeling, radioactivity measurement, CC, cell growth, enzymatic assay, centrifugation, COSY, protein determination
  
   
  
  Rhizobium etli D-1 2002.8
  CSDB ID 47782 (all data & tools)