Outer Membrane Proteins and Lipopolysaccharides in Pathovars of
            <i>Xanthomonas campestris</i>

Ojanen, Tuula; Helander, Ilkka M.; Haahtela, Kielo; Korhonen, Timo; Laakso, Tuula

doi:10.1128/aem.59.12.4143-4151.1993

Cited by 32 publications

(14 citation statements)

References 48 publications

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“…Salmonella OMPs were individually extracted by following the procedure of Ojanen et al (1993). Briefly, the overgrown culture of Salmonella cells was harvested by centrifugation at 15 000 g for 20 min at 4°C.…”

Section: Preparation Of Outer Membrane Protein Extractsmentioning

confidence: 99%

Differentiation of outer membrane proteins from Salmonellaenterica serotypes using Fourier transform infrared spectroscopy and chemometrics

Kim

Reuhs

et al. 2006

Lett Appl Microbiol

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Aims: To differentiate between outer membrane proteins (OMPs) from six Salmonellaenterica serotypes using a Fourier transform infrared (FTIR) spectroscopy method and chemometrics. Methods and Results: The OMPs from Salmonella serotypes (Typhimurium, Enteritidis, Thomasville, Hadar, Seftenberg and Brandenburg) were isolated using a sarcosyl extraction method. OMP profiles on SDS‐PAGE exhibited two or three bands between 48 and 54 kDa. Spectra of 10 μl of OMP preparations (5 mg ml−1) dried on a gold reflective slide were collected using 128 scans at 4 cm−1 resolution and units of log (1/R) and analyzed using canonical variate analysis (CVA) and linear discriminant analysis (LDA). The CVA of Salmonella OMP spectra in the 1800–1500 cm−1 region separated the serotypes and LDA provided a 100% correct classification. Conclusions: The use of a FTIR method combined with chemometrics provided better differentiation of Salmonella OMPs than the OMP pattern analysis by SDS‐PAGE. Significance and Impact of the Study: This is the first study to demonstrate that spectra of OMP extracts from Salmonella serotypes can be used for 100% correct classification of the serotypes studied.

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Section: Preparation Of Outer Membrane Protein Extractsmentioning

confidence: 99%

Differentiation of outer membrane proteins from Salmonellaenterica serotypes using Fourier transform infrared spectroscopy and chemometrics

Kim

Reuhs

et al. 2006

Lett Appl Microbiol

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“…Two other derivatives (NB-9C, program A; R,xl = 0.69 and 0.73, respecsugars, whose alditol acetate derivatives had El mass spectra tively). The L-configuration of these sugars was shown by the typical for those of derivatives of heptoses (29) (37). The identity of the sugar was confirmed when an increased amount of per-O-acetylated rhamnitol was detected after demethylation of sugars of C. fetus LPS with boron tribromide (8), reduction, and then peracetylation.…”

Section: Resultsmentioning

confidence: 88%

“…Covalently bound uronic acid was quantitated by the modified carbazole method of Bitter and Muir (4). Alternatively, uronic acids were characterized and determined by GLC as their carboxyl-reduced (NaB2H4) hexitol acetate derivatives after treatment of LPS by the procedure described previously (37). Analysis of KDO and Neu5Ac.…”

mentioning

confidence: 99%

Biochemical characterization of Campylobacter fetus lipopolysaccharides

et al. 1994

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Lipopolysaccharides (LPS) of five strains of the human and animal pathogen Campylobacter fetus were electrophoretically and chemically characterized. Analysis with sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that all the strains produced smooth-form LPS with 0 side chains of relatively constant chain length. Upon extraction, LPS partitioned into both the water and phenol phases of phenol-water extracts, which showed that two chemical species of LPS were present in each C. fetus strain. Constituents common to all the LPS, though differing in molar ratios, were L-rhamnose, L-fucose, D-mannose, D-glucose, D-galactose, L-glycero-D-manno-heptose, and D-glycero-D-manno-heptose. L-Acofriose (3-0-methyl-L-rhamnose) was present in only two of the C. fetus strains. On the basis of these differences, it was possible to distinguish between LPS from strains of different serotypes and biotypes. Furthermore, chemical analysis indicated that the phenol phase LPS had a lower level of substitution by certain neutral sugars than did water phase LPS. also produce a crystalline, surface array (S-layer) protein (6) which is associated with virulence (5). The S-layer proteins have been demonstrated to attach to the LPS of C. fetus, suggesting that LPS anchors these proteins on the bacterial surface (61). Moreover, inhibition studies with lectins have indicated that the 0 side chain is the region of LPS involved in the interaction (14).Despite the importance of LPS as a toxin and in the serotyping and pathogenesis of C. fetus, no information, apart from comparative immunoblotting data (42,43), is available on the composition and structure of C. fetus high-Mr LPS. This study was undertaken, therefore, to determine the chemical composition of LPS of five C. fetus strains.(A preliminary report of this research was presented at the Seventh International Workshop on Campylobacter, Helicobacter and Related Organisms, 21 to 22 September 1993, Brussels, Belgium). MATERIALS AND METHODSBacterial strains and growth conditions. Five C. fetus strains were used in this study (Table 1). They included strains of both C. fetus subsp. fetus and C. fetus subsp. venerealis and were of different serotypes and biotypes. The strains were maintained at -70°C in tryptone soya broth (Oxoid Ltd., London, England) containing 15% (vol/vol) glycerol and were grown on blood agar (Colombia Agar Base [Oxoid]-7% horse blood) which was incubated in an atmosphere of 5% 02, 10% C02, and 85% N2) at 37°C for 48 h. Salmonella enterica serovar Typhimurium SH2183 was cultivated on L agar as described previously (51). Biomass was harvested in sterile distilled water, centrifuged at 8,000 x g (4°C, 20 min), and washed twice, and the bacterial pellets were freeze dried.Isolation of LPS. To produce minipreparations of LPS, 3922 on July 16, 2020 by guest http://iai.asm.org/ Downloaded from on July 16, 2020 by guest

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“…Both Kdo and sialic acid were quantified as their methyl ketoside methyl ester derivatives by GLC-mass spectrometry (GLC-MS) [7]. Uranic acid was detected by GLC-MS as its deuterium-reduced hexitol acteate as described previously [28] and quantified calorimetrically [16]. Phosphate was assayed calorimetrically [29].…”

Section: Chemical Analytical Methodsmentioning

confidence: 99%

“…Upon high-voltage paper electrophoresis at pH 2.8 the sugar was identified as glucuronic acid by its distinctive mobility CM,,: GlcA, 0.82; galacturonic acid, 0.51; sialic acid, 0.96) and carbazole staining which was comparable with authentic standard (Sigma Chemical Co., St. Louis, MO). Also, hexitol acetates derived from hexuronic acids of LPS which had been subjected to methanolysis and carboxyl reduction by NaBD, were monitored by GLC-MS on the basis of the mass fragment m/z 219 [28]. The only deuterium-reduced hexitol acetate found in samples from each LPS was glucitol acetate.…”

Section: Chemical Analysis Of C Jejuni Lpsmentioning

confidence: 99%

Biological and serological characterization of Campylobacter jejuni lipopolysaccharides with deviating core and lipid A structures

Moran

1995

FEMS Immunology and Medical Microbiology

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Lipopolysaccharides from Campylobacter jejuni were tested for their ability to induce toxic lethality in galactosamine-sensitized mice, pyrogenicity in rabbits and tumour necrosis factor (TNF) secretion from mouse peritoneal macrophages. Compared with those of Salmonella LPS, lethal toxicity was 50% lower, pyrogenicity was 30- to 50-fold lower, and ability to induce TNF was 100-fold lower. C. jejuni LPS and lipid A exhibited higher phase-transition temperatures than those of Salmonella preparations, and thus the former have lower fluidity at 37 degrees C. This lower fluidity of acyl chains may influence the biological activities of C. jejuni LPS, but acyl chain characteristics and diaminoglucose replacing glucosamine in the hydrophilic lipid A backbone may also influence the supramolecular structure of lipid A, thereby affecting biological activities. Although diaminoglucose is present in the backbone of C. jejuni lipid A, antigenically the latter resembled classical lipid A of the Enterobacteriaceae when tested with anti-lipid A antibodies. Chemical investigations suggested the presence of glucuronic acid in an acid labile linkage in the inner core region, thus producing a structurally unusual region in C. jejuni LPS.

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Outer Membrane Proteins and Lipopolysaccharides in Pathovars of Xanthomonas campestris

Cited by 32 publications

References 48 publications

Differentiation of outer membrane proteins from Salmonellaenterica serotypes using Fourier transform infrared spectroscopy and chemometrics

Differentiation of outer membrane proteins from Salmonellaenterica serotypes using Fourier transform infrared spectroscopy and chemometrics

Biochemical characterization of Campylobacter fetus lipopolysaccharides

Biological and serological characterization of Campylobacter jejuni lipopolysaccharides with deviating core and lipid A structures

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