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Query: UNIPROT:P43026 (lipopolysaccharide)
62,215 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A mutant of Escherichia coli that lacks uridine 5'-diphosphate galactose-4-epimerase makes lipopolysaccharide with less carbohydrate than the parent, unless galactose is present during growth. Carbohydrate is dense, and the outer membrane, which contains lipopolysaccharide, was found to be denser when isolated from cells grown with galactose then when galactose was omitted. Cells given galactose after growth in its absence rapidly formed dense regions within the outer membrane that disappeared when galactose was removed. These results indicate that lipopolysaccharide enters the outer membrane nonrandomly at a minimum of 10 to 22 discrete "insertion points." Isopycnic centrifugation provides a method for isolating these regions.
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PMID:Mode of insertion of lipopolysaccharide into the outer membrane of escherichia coli. 77 Apr 32

Several chromosomal loci are involved in lipopolysaccharide (LPS) biosynthesis by Haemophilus influenzae. Two of these, lic1 and lic2, contain multiple open-reading frames (ORFs) and include tandem repeats of the tetramer CAAT within and at the 5' end of the coding region of the first ORF in each locus. Variation in the number of repeats of CAAT is involved in the variable expression of LPS epitopes, and genes within these loci are involved in the biosynthesis of these epitopes. lic3 also contains multiple ORFs and the CAAT repeats in the same arrangement as in the other two lic loci. However, in lic3 metabolic functions are encoded by the downstream genes. ORF2 is galE, encoding uridine 5'-diphosphogalactose 4-epimerase, and ORF4 is adk, encoding adenylate kinase. A mutant H. influenzae with a deleted galE gene had an altered LPS when grown on media lacking galactose and was of reduced virulence in infant rats.
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PMID:Molecular biology of phase-variable lipopolysaccharide biosynthesis by Haemophilus influenzae. 158 88

An Rc-mutant of Escherichia coli that lacks UDPgalactose 4-epimerase grows normally without galactose but makes lipopolysaccharide lacking most of its carbohydrate. Exogenous galactose overrides the mutation and results in the formation of a complete lipopolysaccharide, thereby producing a smooth phenocopy. The smooth phenocopy was much more resistant to the bactericidal activity of normal human serum than was the rough phenotype. More complement was utilized by the rough mutant in the bactericidal process than by the smooth phenocopy. An antiserum was prepared in rabbits to a specific outer membrane protein in the mutant bacterium, the lambda receptor, whose expression could be suppressed by the addition of 10 mM maltose. The effect of the O-antigen in the lipopolysaccharide produced by the smooth phenocopy on the binding of antibody to the lambda receptor was determined. The smooth phenocopy exhibited significantly less binding of antibody than did the rough phenocopy. In addition, expression of the lambda receptor had little effect on the binding of antibody to the lambda receptor in the smooth phenocopy but caused significantly increased binding in the rough mutant. The results suggest that the increased resistance to the lethal action of normal human serum shown by the smooth phenocopy may be due to the blocking of antibody binding sites by the O-antigen of lipopolysaccharide, thereby preventing activation of the classical pathway of complement.
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PMID:Inhibition of serum bactericidal reaction by lipopolysaccharide. 403 5

A class of galactose-resistant mutants has been derived from strains of Salmonella typhimurium which are defective in uridine diphosphoglucose-4-epimerase. Resistant strains are phenotypically similar to parent organisms but do not lyse in the presence of galactose. Low levels of functional epimerase can be detected in induced cells grown at 20 C but not at 37 C, and acid is not produced from galactose. Sufficient galactose is synthesized at reduced temperatures to fabricate smooth lipopolysaccharide and acceptor sites for phage P22 from galactose-deficient media. The leaky nature of these mutants may account for resistance to galactose death by maintaining galactose metabolites at a subcritical level. Glucose protects sensitive strains by control of levels of toxic metabolites by catabolite repression.
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PMID:Resistance of Salmonella typhimurium mutants to galactose death. 489 80

The "T1 side chain" portion of cell wall lipopolysaccharide from T1 strains of Salmonella contains d-galactofuranose and d-ribofuranose residues. Isotope labeling studies, using intact cells of mutants each blocked at either of the two different steps of d-galactose metabolism (uridine diphosphate-glucose 4-epimerase and galactose-1-P uridylyl transferase) or at phosphoglucoisomerase, led to the following conclusions. (i) d-Galactofuranose residues are synthesized from d-galactopyranose or its derivatives, rather than by a direct conversion from other hexopyranoses or their derivatives. (ii) The pyranose-to-furanose conversion does not appear to take place at the level of the free d-galactose or d-galactose 1-phosphate. This result suggests that the conversion may occur at the stage of uridine diphosphate-d-galactose. (iii) In a mutant lacking phosphoglucoisomerase, d-ribofuranose residues in T1 side chains contained (14)C derived from exogenous d-fructose-U-(14)C, but little (3)H from exogenous d-glucose-1-(3)H. Thus, no evidence was found for a direct pathway of aldohexose-to-ribose conversion involving a loss of one of the carbons in the C2-C6 moiety of aldohexoses. This suggests, but does not prove, that the T1 ribofuranose residues are synthesized by conventional mechanisms involving hexose monophosphate shunt and transketolase-transaldolase reactions.
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PMID:Biosynthesis of T1 antigen in Salmonella: origin of D-galactofuranose and D-ribofuranose residues. 492 77

Salmonella typhimurium strains which are deficient in uridine diphosphate (UDP)-galactose-4-epimerase (gal E mutants) owe their outstanding protective capacity when used as live vaccine to the fact that when galactose is supplied exogenously, such as occurs in vivo, smooth cell wall lipopolysaccharides are synthesized. The mutants lose most of their protective capacity when this phenotypic curing is prevented by a second mutation of the kind found in strains LT(2)M(1)A (deficient in galactokinase) or E(32) (deficient in UDP-galactose-lipopolysaccharide transferase). Despite such phenotypic reversion, the gal E mutants are rendered avirulent as a result of galactose-induced bacteriolysis. Secondary mutants have been isolated which differ from each other with respect to the extent of galactose-induced lysis. The differences in galactose sensitivity are attributable to different activities of the other Leoloir pathway enzymes, namely, galactokinase and galactose-1-phosphate-uridyl transferase. The influence of these enzymes on lipopolysaccharide composition and galactose sensitivity and thus on virulence and immunogenicity of gal E mutants has been studied.
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PMID:Immunity in experimental salmonellosis. II. Basis for the avirulence and protective capacity of gal E mutants of Salmonella typhimurium. 494 7

The membrane fraction from a mutant of Salmonella anatum deficient in UDPgalactose-4-epimerase, utilized synthetic ficaprenyl alpha-D-galactosyl diphosphate as a substrate in the biosynthesis of the O-polysaccharide portion of lipopolysaccharide which has a mannosylrhamnosylgalactose repeating sequence. The galactosyl lipid was prepared by chemical synthesis from D-galactose and ficaprenol extracted from Ficus elastica. Membrane preparations catalyzed the transfer of rhamnose from TDP-rhamnose onto membrane-bound ficaprenyl galactosyl diphosphate forming rhamnosylgalactosyl ficaprenyl diphosphate; the reaction was dependent on the prior insertion of the synthetic glycosyl-lipid into the membrane, and was proportional to incubation time up to 4 min at 29 degrees C. When both TDP-rhamnose and GDP-mannose were added, the product formed was O-polysaccharide. These results indicate that the chemically synthesized ficaprenyl galactosyl diphosphate can be an active substrate for the in vitro synthesis of the Salmonella O-polysaccharide.
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PMID:Chemically synthesized galactosyl ficaprenyl diphosphate as an intermediate in the biosynthesis of the Salmonella O-antigenic polysaccharide. 618 14

We studied the relationship between serum antibodies to the cross-reactive endotoxin core of Escherichia coli and survival following Pseudomonas aeruginosa septicemia. Core glycolipid was purified from the outer cell membrane of a uridine diphosphate galactose 4-epimerase-deficient rough mutant E. coli (J5 strain), characterized, and used as the antigen in a quantitative enzyme-linked immunosorbent assay (ELISA) to measure core-specific IgG and IgM antibodies. 43 patients with Pseudomonas septicemia, among whom there was a mortality of 42%, were evaluated. Core-specific antibody concentrations in acute sera ranged from 1 to 49 micrograms/ml in the case of IgG and from 1 to 200 micrograms/ml for IgM. Core-specific antibodies of both isotypes were higher in patients who survived compared with those who succumbed to their septicemias (mean, microgram/ml +/- SEM, 26 +/- 3 vs. 14 +/- 4, P = 0.005 for IgG, and 55 +/- 12 vs. 18 +/- 5, P = 0.009 for IgM). Although total IgG levels were also higher in acute sera from survivors compared with nonsurvivors (mean, mg/dl +/- SEM, 1,120 +/- 99 vs. 694 +/- 119, P = 0.004), total IgM levels were virtually identical in the two groups (146 +/- 23 vs. 148 +/- 48, P = 0.52). Conversely, patients with core-specific IgG levels greater than 10 micrograms/ml at the onset of septicemia had better survival than those with levels less than 10 micrograms/ml (79 vs. 14%, P less than 0.001), and patients with core-specific IgM levels greater than 30 micrograms/ml had better survival than those with levels less than 30 micrograms/ml (81 vs. 44%, P = 0.01). In comparison, patients with total IgG levels greater than 1,000 mg/dl also had better survival than those with levels less than 1,000 mg/dl (82 vs. 42%, P = 0.01), while those with total IgM levels greater than 150 mg/dl showed somewhat less improvement in survival compared with those with levels less than 150 mg/dl (71 vs. 50%, P = 0.12). Core-specific IgM was highly correlated with core-specific IgG (r = 0.52), but not with type-specific anti-lipopolysaccharide (r = 0.13) or anti-toxin A (r = 0.12) antibodies, or with total IgG (r = 0.28) or IgM (r = 0.31). In contrast, core-specific IgG correlated somewhat more closely with type-specific antibodies (r = 0.36), and with total IgG (r = 0.51) and IgM (r = 0.52). Stepwise linear discriminant analysis indicated that type-specific antibody levels were the best predictor of outcome, among those antibodies examined, followed by anti-core IgM. Although anti-core IgG, anti-toxin A, and total IgG levels all correlated individually with survival, none augmented the prognostic power of type-specific antibodies in combination with anti-core IgM, which together predicted outcome accurately 73.5% of the time. Host factors not significantly associated with anti-core antibody levels included rapidly fatal underlying disease, age, sex, leukopenia, and prior treatment with cytotoxic drugs. In contrast, prior steroid therapy was associated with low levels of both core-specific IgG and IgM (P < 0.05). These data suggest cross-protective activity against P. aeruginosa septicemia of naturally occurring antibodies to the endotoxin core of E. coli. Anti-core antibodies, particularly of the IgM isotype appear to augment the more specific protective immunity engendered by antibodies to the O-specific side chains of Pseudomonas lipopolysaccharides. This cross-protective immunity likely applies to other Gram-negative pathogens as well.
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PMID:Enhanced survival in Pseudomonas aeruginosa septicemia associated with high levels of circulating antibody to Escherichia coli endotoxin core. 635 57

Galactose metabolism mutants of Erwinia amylovora were created by transposon insertions and characterized for their growth properties and interaction with plant tissue. The nucleotide sequence of the galE gene was determined. The gene, which encodes UDP-galactose 4-epimerase, shows homology to the galE genes of Escherichia coli, Neisseria gonorrhoeae, Rhizobium meliloti, and other gram-negative bacteria. Cloned DNA with the galE and with the galT and galK genes did not share borders, as judged by the lack of common fragments in hybridization with chromosomal DNA. These genes are thus located separately on the bacterial chromosome. In contrast to the gal operon of E. coli, the galE gene of E. amylovora is constitutively expressed, independently of the presence of galactose in the medium. The function of the galE gene but not of the galT or galK gene is required for bacterial virulence on pear fruits and seedlings. In the absence of galactose, the galE mutant was deficient in amylovoran synthesis. Subsequently, the galE mutant cells elicited host defense reactions, and they were not stained by fluorescein isothiocyanate-labelled lectin, which efficiently binds to amylovoran capsules of E. amylovora. The mutation affected the side chains of bacterial lipopolysaccharide, but an intact O antigen was not required for virulence. This was shown with another mutant, which could be complemented for virulence but not for side chain synthesis of lipopolysaccharide.
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PMID:Genetics of galactose metabolism of Erwinia amylovora and its influence on polysaccharide synthesis and virulence of the fire blight pathogen. 750 2

The inner-core region of the lipopolysaccharide of an UDPGalNAc-4-epimerase-deficient mutant of Yersinia enterocolitica 0:3, designated as Ye75R, was investigated using methylation analysis, 1D-13C-NMR and 2D-13C-NMR and 1H-NMR, as well as 31P-NMR, fast-atom-bombardment mass spectrometry (FAB MS) and FAB MS/MS in positive and negative modes. The isolated core heptasaccharide (OS) was composed of 2 units D-glucose, 3 units LD-heptose and 1 unit each of DD-heptose and 3-deoxy-D-manno-octulosonic acid. Methylation analysis indicated that OS was highly branched with terminal location of the two glucoses and the DD-heptose unit, which was partially (to about 40%) phosphorylated at C7. These combined studies allowed us to formulate the structure of the inner core region as shown in Scheme 1. The substitution of the 7-position of the terminally linked DD-heptose unit by phosphate could be recognized by MS characterization of permethylated DD-heptose-7-phosphate (alditol acetate) and the extent of the substitution by the ratio of the two well separated 1H signals of DD-heptose in 500-MHz 1H-NMR. Negative FAB MS of OS also indicated the presence of smaller amounts of two hexasaccharides, differing from OS in lacking either one terminal unit of D-glucose or of the terminal DD-heptose, and additionally of a pentasaccharide lacking two heptosyl units, namely the terminal DD-heptose and and the subterminal LD-heptose. The presence of the smaller oligosaccharides in the OS fraction was also recognized by the methylation analysis.
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PMID:The inner core region of Yersinia enterocolitica Ye75R (0:3) lipopolysaccharide. 816 22


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