Structural and immunochemical characterization of the O-haptens of Brucella abortus lipopolysaccharides from strains 19 and 2308 |
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| Authors: | Albert M. Wul Neil E. Mackenzie |
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| Affiliation: | (1) Department of Veterinary Pathology College of Veterinary Medicine, Texas Agricultural Experiment Station, Texas A&M University, 77843 College Station, Texas, USA;(2) Departments of Chemistry and Veterinary Microbiology and Parasitology, College of Veterinary Medicine, Texas Agricultural Experiment Station, Texas A&M University, 77843 College Station, Texas, USA;(3) Present address: The Upjohn Co., 7000 Portage Road, 7837-259-12, 49001 Kalamazoo, MI, USA |
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| Abstract: | The O-haptens of the major fraction (f5A) of B. abortus (Strains 2308 and 19) membrane bound smooth lipopolysaccharide (sLPS) were prepared by hydrolysis of f5A native sLPS in 1% acetic acid at 100°C for 2 h. After hydrolysis, O-haptens were separated from Lipid A-protein complex by centrifugation, and from small fragments by ultrafiltration of molecular weight cut-off (MWCO) 1.0 × 103. These carbohydrate haptens were identified by precipitin-inhibition assay and further fractionated by both membrane filtration and dialysis. The size distributions of carbohydrate haptens of endotoxins (f5A) ranged from oligosaccharides up to polysacchandes of 1.0 × 104 MWCO. Three major fractions of MWCO 8.0 – 10.0 × 103, 3.5 – 5.0 × 103 and < 1.0 × 103 from both strains 2308 and 19 contained more than 85% of the total immunoactive materials. These fractions of haptens were subjected to composition, proton and 13C NMR analysis and were found to be a homopolymer of  1 2 linked, 4,5-dideoxy-4-formamido-D-mannose (N-formylperosamine), which is identical to O-haptens of B. abortus strain 119.3 and Yersinia enterocolitica serotype 0:9 and similar to Vibrio cholerae 569B (INABA).Fractions of these haptens exhibited similar inhibitory reactivities in a precipitin-inhibition assay as expressed as µmoles of monosaccharide of anhydro-N-formyl perosamine. They were about 480 times as active as Me DMan or DMan.Abbreviations LPS lipopolysaccharides - sLPS smooth lipopolysaccharides - cLPS crude lipopolysaccharide(s) which is equivalent to sLPS of f5, prepared by Moreno et al. [8] - f5A fraction A of f5 which is one of the major of crude LPS prepared by the modification of the method of Moreno et al. [5] - fr. or Fr. fraction - dH2O distilled H2O - AH acid hapten [20] - KDO 3-deoxyoctuosonic acid - DGIc DGlucopyranose - DGal DGalactopyranose - DMan DMannopyranose - MeDMan Methyl-DMan-nopyranoside - MWCO Molecular weight cut off |
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| Keywords: | O-haptens lipopolysaccharides Brucella abortus |
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