Isolation and characterization of the lipopolysaccharides from Bradyrhizobium japonicum

Carrion, Miguel; Bhat, U. Ramadas; Reuhs, Bradley L.; Carlson, Russell W.

doi:10.1128/jb.172.4.1725-1731.1990

Cited by 48 publications

(38 citation statements)

References 45 publications

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“…Galacturonic acid has so far not been reported to occur in lipid A samples from any gram-negative bacteria. The type 3 lipid A's contain either only DAG or glucosamine and DAG (mixed lipid A backbone), as has already been reported for B. japonicum (11,24) and Bradyrhizobium sp. (Lupinus) strain DSM 30140 (24).…”

Section: Methodssupporting

confidence: 62%

Occurrence of lipid A variants with 27-hydroxyoctacosanoic acid in lipopolysaccharides from members of the family Rhizobiaceae

et al. 1991

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Lipopolysaccharides (LPSs) isolated from several strains of Rhizobium, Bradyrhizobium, Agrobacterium, and Azorhizobium were screened for the presence of 27-hydroxyoctacosanoic acid. The LPSs from all strains, with the exception of Azorhizobium caulinodans, contained various amounts of this long-chain hydroxy fatty acid in the lipid A fractions. Analysis of the lipid A sugars revealed three types of backbones: those containing glucosamine (as found in Rhizobium meliloti and RhizobiumfrediO), those containing glucosamine and galacturonic acid (as found in Rhizobium leguminosarum bv. phaseoli, trifolii, and viciae), and those containing 2,3-diamino-2,3-dideoxyglucose either alone or in combination with glucosamine (as found in Bradyrhizobium japonicum and Bradyrhizobium sp.[Lupinus] strain DSM 30140). The distribution of 27-hydroxyoctacosanoic acid as well as analysis of lipid A backbone sugars revealed the taxonomic relatedness of various strains of the Rhizobiaceae.Bacteria belonging to the family Rhizobiaceae are gram negative and are able to form nitrogen-fixing symbiotic relationships with legume plants. There are three distinct genera: the symbiotic nitrogen-fixing Rhizobium and Bradyrhizobium spp. and the plant pathogenic Agrobacterium spp. Quite recently a new genus, so far comprising only the stem-nodulating nitrogen-fixing species Azorhizobium caulinodans, was defined (13). Of these genera, the species of Rhizobium are taxonomically closely related and show genetic similarities to the genus Agrobacterium as evidenced by 16S rRNA homology studies (12). On the other hand, the slowly growing species of Bradyrhizobium are rather distantly related to the other two genera as revealed by their low SAB values determined by DNA-rRNA hybridization studies (1). In addition to the nucleotide sequence homology studies, differentiation of various members of the Rhizobiaceae has been attempted by several chemotaxonomic approaches such as cellular fatty acid analysis (21, 31), polyacrylamide gel electrophoresis of cellular proteins (19), and composition analysis of extracellular gum (26). However, results of these studies were not sufficient to adequately distinguish between members of the Rhizobiaceae. More recently, the backbone sugar composition of lipid A fractions of lipopolysaccharide (LPS) has been used as a taxonomic marker for recognition and relatedness of various nonsulfur bacteria (23). Therefore, in this study, the lipid A fractions from rhizobial LPSs were examined to see whether they represented a marker for determining the relatedness of these bacteria.The surface polysaccharides, including the LPS, of strains of Rhizobium have been hypothesized to be involved in the molecular mechanisms of symbiotic infection (5). In an attempt to elucidate the structure of LPS from rhizobial strains, an unusual very-long-chain hydroxy fatty acid, 27-hydroxyoctacosanoic acid (27-OH-28:0), was discovered to be the major fatty acid constituent of the lipid A region (15). More recently, we have also identified this long-cha...

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Section: Methodssupporting

confidence: 62%

Occurrence of lipid A variants with 27-hydroxyoctacosanoic acid in lipopolysaccharides from members of the family Rhizobiaceae

et al. 1991

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“…All Rhizobiaceae (i.e. the plant symbiontic rhizobia) and a few other Rhizobiales, including intracellular mammalian pathogens Bartonella and Brucella, examined to date contain a ( -1)-hydroxylated long-chain fatty acids in their lipid A, likely reflecting their phylogenetic relationship (11,29). However, be- cause of a limited number of detailed structural studies on lipid A's from these organisms, and because of difficulties associated with analyzing the long-chain fatty acids, the location, stoichiometry, and type of attachment of these substituents is known only for a few species, including Sinorhizobium sp.…”

Section: Discussionmentioning

confidence: 99%

“…The lipid A backbone of B. henselae ATCC 49882 T is composed of a bisphosphorylated GlcN3N disaccharide shared with only a few bacteria, including Pseudomonas diminuta, Bradyrhizobium japonicum (29), and L. pneumophila (32). Lipid A of Campylobacter jejuni, which is thus far the most thoroughly investigated representative of a GlcN3N-containing lipid A, is composed predominately of a hybrid GlcN3N-GlcN disaccharide, whereas GlcN3N-GlcN3N is found in minor LPS species (24).…”

Section: Discussionmentioning

confidence: 99%

Structure and Biological Activity of the Short-chain Lipopolysaccharide from Bartonella henselae ATCC 49882T

Zähringer

Lindner

Knirel

et al. 2004

Journal of Biological Chemistry

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The facultative intracellular pathogen Bartonella henselae is responsible for a broad range of clinical manifestations, including the formation of vascular tumors as a result of increased proliferation and survival of colonized endothelial cells. This remarkable interaction with endotoxin-sensitive endothelial cells and the apparent lack of septic shock are considered to be due to a reduced endotoxic activity of the B. henselae lipopolysaccharide. Here, we show that B. henselae ATCC 49882 T produces a deep-rough-type lipopolysaccharide devoid of O-chain and report on its complete structure and Toll-like receptordependent biological activity. The major short-chain lipopolysaccharide was studied by chemical analyses, electrospray ionization, and matrix-assisted laser desorption/ ionization mass spectrometry, as well as by NMR spectroscopy after alkaline deacylation. The carbohydrate portion of the lipopolysaccharide consists of a branched trisaccharide containing a glucose residue attached to position 5 of an ␣-(234)-linked 3-deoxy-Dmanno-oct-2-ulosonic acid disaccharide. Lipid A is a pentaacylated ␤-(1 36)-linked 2,3-diamino-2,3-dideoxyglucose disaccharide 1,4 -bisphosphate with two amidelinked residues each of 3-hydroxydodecanoic and 3-hydroxyhexadecanoic acids and one residue of either 25-hydroxyhexacosanoic or 27-hydroxyoctacosanoic acid that is O-linked to the acyl group at position 2 . The lipopolysaccharide studied activated Toll-like receptor 4 signaling only to a low extent (1,000 -10,000-fold lower compared with that of Salmonella enterica sv. Friedenau) and did not activate Toll-like receptor 2. Some unusual structural features of the B. henselae lipopolysaccharide, including the presence of a long-chain fatty acid, which are shared by the lipopolysaccharides of other bacteria causing chronic intracellular infections (e.g. Legionella and Chlamydia), may provide the molecular basis for low endotoxic potency.

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“…The water phase containing the LPS was treated with RNase, DNase, and proteinase K, dialyzed, and then lyophilized (22). The LPS extracted into the phenol phase was treated as described by Carrion et al (23). The LPS was purified from these crude preparations with affinity chromatography using polymyxin B-Sepharose (Pierce) (24,25).…”

Section: Methodsmentioning

confidence: 99%

Rhizobium etli CE3 Bacteroid Lipopolysaccharides Are Structurally Similar but Not Identical to Those Produced by Cultured CE3 Bacteria

D’Haeze

Leoff

Freshour

et al. 2007

Journal of Biological Chemistry

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Root nodule development is orchestrated by a symbiotic molecular dialogue between Gram-negative Rhizobium bacteria (e.g. Azorhizobium sp., Bradyrhizobium sp., Rhizobium sp., Sinorhizobium sp.) and specific legume host plants. Nodules are newly formed organs consisting of plant cells occupied with bacteroids that provide the host plant with fixed nitrogen. In the best studied symbiotic interactions, bacteria enter the roots via susceptible curled root hairs, and intracellular infection threads guide the bacteria toward de novo nodule primordia, where internalization into plant cells takes place. Initiation of nodule development and invasion require the production of bacterial signal molecules, including fatty acylated chitin oligosaccharides known as Nod factors (1), and structurally complex surface polysaccharides (SPS) 3 (2, 3). The outer surface of rhizobia typically consists of SPS that include extracellular polysaccharides (EPS) that are released into the media, capsular polysaccharides that are tightly associated with the bacterial surface, and lipopolysaccharides (LPS) that are anchored in the outer membrane (4). LPS are composed of lipid A, a core oligosaccharide, and an O-antigen polysaccharide. Accumulating data demonstrate the important role that rhizobial SPS play in invasion and nodule development and their involvement in the initiation of infection and invasion, suppression of plant defense, bacterial release from infection threads, bacteroid development and senescence, induction of plant gene expression, and protection against antimicrobial compounds (2, 3).Various observations suggest that proper LPS synthesis is required for invasion and nodule development in various symbiotic interactions, including the interaction between Rhizobium etli and Phaseolus vulgaris (2, 4). An R. etli mutant that lacks the O-chain polysaccharide portion of its LPS elicited the formation of infection threads on P. vulgaris; however, the bacteria ceased to develop within the root hair that formed thick walls (5, 6). The formation of nodule primordia was normal, but no bacteria were released from infection threads and internalized into plant cells (6). Occasionally, some bacteria were present in intercellular spaces. It was furthermore demonstrated that not only the presence of the O-chain polysaccharide on the LPS but also the abundance of O-chain polysaccharide was important for nodulation. For example, mutant strain R. etli CE166 produced, based on PAGE analysis of the LPS, only 40% LPS containing the O-chain polysaccharide compared with the parent strain, and the symbiotic phenotype of this mutant was

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Isolation and characterization of the lipopolysaccharides from Bradyrhizobium japonicum

Cited by 48 publications

References 45 publications

Occurrence of lipid A variants with 27-hydroxyoctacosanoic acid in lipopolysaccharides from members of the family Rhizobiaceae

Occurrence of lipid A variants with 27-hydroxyoctacosanoic acid in lipopolysaccharides from members of the family Rhizobiaceae

Structure and Biological Activity of the Short-chain Lipopolysaccharide from Bartonella henselae ATCC 49882T

Rhizobium etli CE3 Bacteroid Lipopolysaccharides Are Structurally Similar but Not Identical to Those Produced by Cultured CE3 Bacteria

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