Biomedical subjects
Publications and source records attributed to L A Parolis.
The primary structure of the O-antigen of Escherichia coli O117 was shown by monosaccharide analysis, methylation analysis, and by 1D and 2D 1H and 13C NMR spectroscopy to be composed of linear pentasaccharide repeating units with the structure: -->3)-alpha-D-GalpNAc-(1-->4)-beta-D-GalpNAc-(1-->3)-alpha-L-Rhap- (1-->4)- alpha-D-Glcp-(1-->4)-beta-D-Galp-(1-->
The primary structure of the O-antigen of Escherichia coli O116:K+:H10 was shown by monosaccharide analysis, a partial hydrolysis study and by 1D and 2D 1H and 13C NMR spectroscopy to be composed of linear pentasaccharide repeating units with the structure: -->6)-alpha-D-GlcpNAc-(1-->4)-alpha-D-GalpNAc-(1-->4)-alpha-D-GalpA++ +-(1-->3)- beta-D-GlcpNAc-(1-->2)-beta-D-Quip4NAc-(1-->.
The structure of a linear, acidic exopolysaccharide isolated from the Archaeon Haloferax denitrificans ATCC 35960 has been determined using NMR spectroscopy. The sugar residues in the repeating unit of the polysaccharide were identified as Gal and GlcA2,3NAc after the assignment of the 1H and 13C resonances using COSY, HOHAHA, HMQC and HMQC-TOCSY experiments. The sequence of the residues in the polysaccharide was established from the inter-residue connectivities observed in the HMQC-NOESY plot. The only sugar released on acid hydrolysis was shown to be D-Gal by GLC analysis, while the absolute configuration of the acidic sugars was shown to be D by comparison of the carbon chemical shifts with those of model compounds. Partial acid hydrolysis yielded a tetrasaccharide, terminated by D-Gal at the reducing end, whose structure confirmed that of the repeating unit of the polysaccharide as-->4)-beta-D-GlcpA2,3NAc-(1-->4)-beta-D-GlcpA2, 3NAc-(1-->4)-alpha-D-GlcpA2,3NAc-(1-->3)-alpha-D-Galp- (1-->, where D-GlcpA2,3NAc is 2,3-diacetamido-2,3-dideoxy-D-glucopyranosiduronic acid.
The exopolysaccharide produced by Pichia (Hansenula) holstii NRRL Y-2448 is composed of a phosphomannan core to which oligosaccharide diester phosphate side chains are appended. The oligosaccharides of the side chains were released as oligosaccharide phosphates and neutral oligosaccharides by mild hydrolysis with aqueous acetic acid and aqueous hydrogen fluoride, respectively. The liberated oligosaccharide phosphates were studied by NMR spectroscopy and by electrospray and fast atom bombardment mass spectrometry. The structures of the neutral oligosaccharides were determined by 1D and 2D NMR spectroscopic experiments. Further insight into the length of the side chains was obtained from a matrix assisted laser desorption ionisation-time of flight mass spectrometric study of high and low molecular weight fragments obtained from partial acid hydrolysis of the native polysaccharide.
The structure of the neutral exocellular polysaccharide isolated from the Archaeon Haloferax gibbonsii (ATCC 33959) has been determined using acid hydrolysis, methylation analysis and NMR spectroscopy. The polysaccharide contained D-Man, D-Glc, D-Gal and L-Rha in the ratios 2:1:3:1. The substitution patterns of the sugar residues were deduced from the methylation analysis which indicated the polymer to be composed of a heptasaccharide repeating unit containing two branches. The 1H and 13C NMR resonances of the component sugars were assigned using COSY, HOHAHA, HMQC, and HMQC-TOCSY 2D NMR experiments and the sequence of the sugars in the repeating unit was determined from NOESY and HMBC experiments. The structure can be written as: [formula: see text]
The primary structure of the acidic capsular antigen of Escherichia coli O9:K33:H- was shown by glycose analysis, methylation analysis, and by 1D and 2D 1H and 13C NMR spectroscopy to be composed of branched tetrasaccharide repeating units with the structure: [formula: see text]
The O-specific polysaccharide isolated from the lipopolysaccharide (LPS) of Escherichia coli O121 by mild acid hydrolysis has been studied using mainly NMR spectroscopy. The polysaccharide was treated with mild base to yield a O-deacetylated polysaccharide which contained D-GlcNAc, D-GalNAcA, D-GalNAcAN (2-acetamido-2-deoxy-D-galacturonamide) and D-Qui4NAcGly (where D-Qui4N is 4-amino-4,6-dideoxy-D-glucose) in equimolar proportions. The presence of the amide was confirmed by recording the 1H NMR spectrum of the O-deacetylated polysaccharide at different pH values. The O-acetyl group was located on O-3 of the GalNAcAN and the structure of the polysaccharide can be written as [sequence: see text] This structure is almost identical to that previously reported for the O-specific polysaccharide of Shigella dysenteriae type 7 LPS, the only difference being that O-acetylation is stoichiometric in the latter.
The primary structure of the acidic capsular antigen of Escherichia coli O8:K41:H11 was shown by monosaccharide analysis, methylation analysis, and by 1D and 2D (1)H amd (13)C NMR spectroscopy to be composed of branched pentasaccharide repeating units with the structure (formula: see text)
The halophilic Archaeon Haloferax mediterranei exudes into the growth medium a high molecular weight sulfated polysaccharide. The structure of the repeating unit of this polymer was determined by a combination of glycose, methylation, and sulfate analysis, periodate oxidation, and 1D and 2D NMR spectroscopic analysis of the native and periodate-oxidised/reduced polysaccharides. The location of the sulfate group was established from the 1H and 13C NMR data. The structure of the repeating unit of the polysaccharide may be written as [formula: see text]
The phosphomannan core of the exopolysaccharide of Pichia (Hansenula) holstii NRRL Y-2448 was isolated after hydrolytic removal of the oligosaccharide phosphate side-chains. The core polysaccharide and its dephosphorylated derivative were subjected to extensive 1D and 2D NMR spectroscopy which yielded information on the linkage sites and on the sequence of the mannosyl residues in the major oligosaccharide repeating unit. The most probable structure for the repeating unit was -[6-O-PO3H2-alpha-D-Man-(1-->3)-alpha-D-Man-(1-->2)-alpha-D-Man-(1 -->2)]-alpha-D-Man-(1-->6)-[alpha-D-Man-(1-->2)]-alpha-D-Man-(1-->6)-. A semiquantitative conformational analysis was performed by Monte Carlo simulations and the result was confirmed by comparison with the experimentally determined NMR data. The distance distribution for the phosphate groups was determined from the modeling and was found to cover the expected range of distances for phosphorylated high-mannose oligosaccharides.
The primary structure of the acidic capsular polysaccharide of Escherichia coli 08:K43:H11 was shown by monosaccharide analysis, methylation analysis, beta-elimination, and by 1D and 2D 1H and 13C NMR spectroscopy to be composed of branched pentasaccharide repeating units with the structure.
The O-specific polysaccharide from Escherichia coli O113 lipopolysaccharide was separated from the core and lipid A by mild acid hydrolysis and purified by GPC. Methylation analysis and 1H and 13C NMR spectroscopic studies of the O-deacetylated polysaccharide allowed the determination of the structure of the pentasaccharide repeating unit of the polysaccharide which can be written as [equation: see text] The position of the O-acetyl groups was not determined.
Klebsiella pneumoniae strains of the K2 capsular serotype are usually highly virulent in mice, which is in contrast to the low virulence of most other serotypes. Here we used a genetic approach to examine the relative contribution of capsule type to the virulence of K. pneumoniae in mice. We used wild-type strains expressing capsular polysaccharide (CPS) serotypes K2 (strain KPA1) and K21a (strains KPB1 and KPC1), which were then used to construct capsule-switched derivatives. The close proximity of the cps gene cluster to selectable his markers made it possible to mobilize the cps genes by conjugation from one serotype (donor) to another (recipient) and to obtain recombinants in which interserotype switching had occurred by reciprocal recombination. Each capsule-switched derivative examined of the KPA and KPC strain backgrounds produced a CPS that was immunologically and structurally identical to that of the donor. Strain background was confirmed by demonstrating restriction fragment length polymorphism patterns identical to those of the respective recipients. The parent strains were then compared with capsule-switched recombinants for phenotypic properties associated with virulence. Clearance from the bloodstreams of mice was rapid in serotype K21a strains of either wild-type or recombinant origin, whereas K2 strains remained viable in the blood during the period examined. These differences appeared to be dependent upon the CPS type but independent of strain background. Binding to macrophages was higher in K21a strains than in those with the K2 capsule and was also independent of the strain background. Both blood clearance and macrophage-binding activities were completely inhibited by yeast mannan, suggesting that they were mediated via the macrophage mannose receptor. The K2 parent strain was highly virulent to mice (50% lethal dose [LD50], 3 x 10(3)), while the K21a parent strains demonstrated low virulence (LD50, > 2 x 10(8)). Interestingly, the virulence of recombinant KPC10(cpsK2), originally of the KPC1(cpsK21a) background, was intermediate (LD50, 4 x 10(5)). In contrast, both cpsK21a recombinants of the originally virulent KPA1 (cpsK2) background became nearly avirulent (LD50, > 2 x 10(8)). Six additional serotypes (K12, K24, K32, K55, K62, and K67) were examined, and all showed a positive correlation between the ability of the Klebsiella serotype to interact with a human mannose receptor, as expressed by Cos I cell recombinants, and the LD50 of the serotype. These results suggest that expression of a capsule which is recognized by the mannose receptor markedly affects the interaction with macrophages and blood clearance.(ABSTRACT TRUNCATED AT 400 WORDS)
The primary structure of the acidic capsular antigen of Escherichia coli O8:K25:H9 was shown by monosaccharide analysis, methylation analysis, and by 1D and 2D 1H and 13C NMR spectroscopy to be composed of linear tetrasaccharide repeating units having the structure:
NMR spectroscopy was performed on the depyruvated capsular antigen of E. coli K103 and on the oligosaccharide obtained by depolymerisation of the native polysaccharide with a viral-borne endoglycanase. This capsular polysaccharide is the only one to be co-expressed with O group 101 and joins a small group of unusual capsular polysaccharides which possess pyruvic acid as the only acidic function. The primary structure was shown to be composed of the repeating unit: [formula: see text]
The primary structure of the acidic capsular antigen produced by E. coli K84 was shown by glycose and methylation analysis, and by 1D and 2D 1H and 13C NMR studies of the polysaccharide and an oligosaccharide produced by lithium-ethylenediamine degradation of the polysaccharide, to be comprised of linear hexasaccharide repeating units of the following structure: -->4)alpha-D-GaI pA-(1-->3)-alpha-D-Man p-(1-->4)-beta-D-GIc p-(1-->3)-alpha- D-Glc pNAc-(1-->2)-beta-D-Man p-(1-->3)-beta-D-Man pNAc-(1-->.
The primary structure of the acidic capsular antigen of Escherichia coli O8:K50:H- was shown by glycose analysis, methylation analysis, and one- and two-dimensional 1H and 13C NMR spectroscopy to be composed of repeating linear tetrasaccharide units having the structure: (formula: see text).
The structure of the exocellular capsular polysaccharide expressed by Escherichia coli O8:K48:H9 has been investigated by hydrolysis and methylation analysis, and by NMR spectroscopic studies of the polysaccharide and of the oligosaccharides generated by solvolysis with anhydrous HF. The capsular polysaccharide was shown to have the repeating unit: [formula: see text] where beta-L-Sug p represents 2,3-diacetamido-2,3,6-trideoxy-beta-L- mannopyranose.