Identification of phosphorylated 3-deoxy-<i>manno</i>-octulosonic acid as a component of <i>Haemophilus influenzae</i> lipopolysaccharide

Zamze, Susanne; Ferguson, Michael A. J.; Moxon, E. Richard; Dwek, Raymond A.; Rademacher, Thomas W.

doi:10.1042/bj2450583

Cited by 46 publications

(27 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…If KDO is detected in the extracellular culture fluid, respcctively. Activity phosphorylated, it does not react efficiently in the thiobarbituof the chloride-stimulated cellobiosidase was mainly found in ric acid assay (61), but because of obviously interfering subthe extracellular culture fluid. The ratio of the enzyme activity stances, this marker was not pursued further.…”

Section: Resultsmentioning

confidence: 99%

Separation of outer and cytoplasmic membranes of Fibrobacter succinogenes and membrane and glycogen granule locations of glycanases and cellobiase

Gong

Forsberg

1993

J Bacteriol

View full text Add to dashboard Cite

The outer membrane (OM) of Fibrobacter succinogenes was isolated by a combination of salt, sucrose, and water washes from whole cells grown on either glucose or cellulose. The cytoplasmic membrane (CM) was isolated from OM-depleted cells after disruption with a French press. The OM and membrane vesicles isolated from the extracellular culture fluid of cellulose-grown cells had a higher density, much lower succinate dehydrogenase activity, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profiles different from those of the CM. The OM from both glucose-and cellulose-grown cells and the extracellular membrane vesicles from cellulose-grown cultures exhibited higher endoglucanase, xylanase, and acetylesterase activities than the CM and other cell fractions. Endoglucanase 2 was absent from the isolated OM fractions of glucose-and cellulose-grown cells and from the extracellular membrane vesicles of cellulose-grown cells but was present in the CM and intracellular glycogen granule fractions, while endoglucanase 3 was enriched in the OM. Cellobiosidase was located primarily in the periplasm as previously reported, while cellobiase was mainly present in the glycogen granule fraction of glucose-grown cells and in a nongranular glycogen and CM complex in cellulose-grown cells. The cellobiase was not eluted from glycogen granules by cellobiose, maltose, and maltotriose nor from either the granules or the cell membranes by nondenaturing detergents but was eluted from both glycogen granules and cell membranes by high concentrations of salts. The eluted cellobiase rebound almost quantitatively when diluted and mixed with purified glycogen granules but exhibited a low affinity for Avicel cellulose. Thus, we have documented a method for isolation of OM from F. succinogenes, identified the OM origin of the extracellular membrane vesicles, and located glycanases and cellobiase in membrane and glycogen fractions.Fibrobacter succinogenes S85 is one of the major cellulolytic bacteria in the rumen of animals receiving low-quality forage rations (44). It extensively degrades cellulose after attachment to the insoluble substrate (39), which indicates that the cellulase enzymes are functioning at the cell surface during cellulose digestion. Nevertheless, the mechanism by which the bacterium degrades cellulose is still unknown (18,44). To elucidate the physiological mechanism of cellulose degradation by the bacterium, three endoglucanases, endoglucanase 1 (EGI), EG2 (46), and EG3 (49), a chloride-stimulated cellobiosidase (31), and a cellodextrinase (28, 29) were purified and characterized. EG I was released from cells during growth, and

show abstract

Section: Resultsmentioning

confidence: 99%

Separation of outer and cytoplasmic membranes of Fibrobacter succinogenes and membrane and glycogen granule locations of glycanases and cellobiase

Gong

Forsberg

1993

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…Phosphorylated sugars were determined indirectly by recording the increase of GC peak area after dephosphorylation of samples with aqueous 48 YO (w/w) hydrofluoric acid at 4 "C for 72 h (Sonesson e t al., 1989a). Phosphorylated sugars were also detected by HPAEC-PED, ASA and by GC analysis of trimethylsilyl (TMS) derivatized methyl glycosides (Zamze et al , 1987). Absolute configurations of the sugars were determined by GC analysis of 2-(R)-butyl and 2-(S)-butyl glycoside diastereomeric derivatives (Gerwig e t al., 1978) after TFA derivatization and retention time comparison to authentic standards.…”

Section: Methodsmentioning

confidence: 99%

Chemical characterization of lipopolysaccharides from Legionella feeleii, Legionella hackeliae and Legionella jordanis

Sonesson¹,

Jantzen²,

Tangen³

et al. 1994

Microbiology

View full text Add to dashboard Cite

Lipopolysaccharides (LPS) from Legionella feeleii serogroup 1 , L. hackeliae serogroup 1 and L. jordanis were subjected to chemical analysis. All three LPS contained D-mannose, D-gIUCOSe, D-glucosamine, L-glycero-D-manno-heptose, 2-keto-3-deoxyoctonic acid and glycerol. In addition the LPS of L. feeleii was characterized by L-quinovose (tentatively identified) and L-fucosamine, L. hackeliae LPS by D-quinovosamine, D-galactosamine and D-galacturonic acid, and L. jordanis LPS by D-quinovosamine. Phosphorylated sugars were detected in all three LPS. The backbone sugar of the lipid A part was in each case 283-diamino-2,3-dideoxy-~-glucose substituted with a complex pattern of fatty acid, including 20-22 different amide-linked (non-branched and methylbranched) 3-hydroxy fatty acids of chain-length ranging from 12 to 23 carbon atoms. The fatty acid patterns included also ester-linked nonhydroxylated entities and the uncommon 27-oxo-octacosanoic acid and 29-oxotriacontanoic acid. The LPS of L. hackeliae and L. jordanis also contained heptacosane-1,27-dioic and nonacosane-1,29-dioic acid, and their 2-hydroxy analogues were characteristic of L. jordanis LPS. SDS-PAGE patterns of the three LPS were distinctly different. Both L. feeleii and L. jordanis produced smooth-form LPS with characteristic ladder patterns, whereas L. hackeliae LPS were of more rough-type character.

show abstract

“…þ and the wild-type strain RM7004 were cultivated as described by Zamze et al (1987). Bacteria were washed with ethanol, acetone (twice), and ether, and dried.…”

Section: Bacteria and Bacterial Lps H Influenzae I-69 Rdmentioning

confidence: 99%

“…influenzae LPS consists of an oligosaccharide (Phillips et al, 1993), with a general architecture similar to that in the intensively studied enterobacteria (Holst and Brade, 1992), linked to the lipid A moiety via a phosphorylated 3-deoxy-D-manno-octulopyranosonic acid (Kdo) residue. Studies on the H. influenzae mutant strain I-69 Rd ¹ /b þ (hereafter referred to as I-69) revealed that it contained a truncated LPS composed of lipid A and a single phosphorylated Kdo residue (Zamze et al, 1987). I-69 was identified from a genomic library of chromosomal DNA in a lambda vector when it was observed that transformants using DNA from one of the lambda clones produced an opaque phenotype (Zwahlen et al, 1985).…”

Section: Introductionmentioning

confidence: 99%

Characterization of monoclonal antibodies recognizing three distinct, phosphorylated carbohydrate epitopes in the lipopolysaccharide of the deep rough mutant I‐69 Rd⁻/b⁺ of Haemophilus influenzae

Różalski

Brade

Kosma

et al. 1997

Molecular Microbiology

View full text Add to dashboard Cite

Summary Monoclonal antibodies against the lipopolysaccharide (LPS) of the deep rough mutant I-69 Rd¹ /b þ of Haemophilus influenzae were obtained after immunization of mice with sheep erythrocytes which had been coated with de-O-acylated LPS. Characterization of antibodies was performed by enzyme immuno assay (EIA) using LPS or neoglycoconjugates containing partial structures of LPS as solid-phase antigens and by haemagglutination with sheep erythrocytes coated with de-O-acylated LPS. Binding data were confirmed by EIA inhibition experiments using deacylated LPS or synthetic partial structures thereof. Three antibodies were specific for 3-deoxy-D-manno -octulopyranosonic acid-(Kdo) 5-phosphate, one for Kdo-4-phosphate, and one required, in addition to a Kdo-phosphate, parts of the phosphorylated glucosamine backbone of lipid A. All antibodies also bound in (i) Western blots to bacterial whole-cell lysates or isolated LPS separated by SDS-PAGE, (ii) bacterial colony blots, and (iii) immunofluorescence with live bacteria. The latter result indicated that Kdo-4-and Kdo-5-phosphate are synthesized by the bacteria and are not the result of phosphate migration.

show abstract

Identification of phosphorylated 3-deoxy-manno-octulosonic acid as a component of Haemophilus influenzae lipopolysaccharide

Cited by 46 publications

References 22 publications

Separation of outer and cytoplasmic membranes of Fibrobacter succinogenes and membrane and glycogen granule locations of glycanases and cellobiase

Separation of outer and cytoplasmic membranes of Fibrobacter succinogenes and membrane and glycogen granule locations of glycanases and cellobiase

Chemical characterization of lipopolysaccharides from Legionella feeleii, Legionella hackeliae and Legionella jordanis

Characterization of monoclonal antibodies recognizing three distinct, phosphorylated carbohydrate epitopes in the lipopolysaccharide of the deep rough mutant I‐69 Rd⁻/b⁺ of Haemophilus influenzae

Contact Info

Product

Resources

About

Identification of phosphorylated 3-deoxy-manno-octulosonic acid as a component of Haemophilus influenzae lipopolysaccharide

Cited by 46 publications

References 22 publications

Separation of outer and cytoplasmic membranes of Fibrobacter succinogenes and membrane and glycogen granule locations of glycanases and cellobiase

Separation of outer and cytoplasmic membranes of Fibrobacter succinogenes and membrane and glycogen granule locations of glycanases and cellobiase

Chemical characterization of lipopolysaccharides from Legionella feeleii, Legionella hackeliae and Legionella jordanis

Characterization of monoclonal antibodies recognizing three distinct, phosphorylated carbohydrate epitopes in the lipopolysaccharide of the deep rough mutant I‐69 Rd−/b+ of Haemophilus influenzae

Contact Info

Product

Resources

About

Characterization of monoclonal antibodies recognizing three distinct, phosphorylated carbohydrate epitopes in the lipopolysaccharide of the deep rough mutant I‐69 Rd⁻/b⁺ of Haemophilus influenzae