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A K Bhattacharjee

Publications and source records attributed to A K Bhattacharjee.

At least 55 records · Page 3Linked to original sources

Purification and characterization of H.8 antigen from group B Neisseria meningitidis.

The surface antigen (H.8) common to the pathogenic Neisseria species was purified by a simple procedure by use of high-performance liquid chromatography. The purified H.8 antigen was characterized as to its amino acid composition, susceptibilities to several proteolytic enzymes, isoelectric point, and susceptibilities to an acid and a base. The amino acid composition of purified H.8 antigen from two strains of Neisseria meningitidis group B, namely, 44/76 and 8047, were compared. It was found that glutamic acid, alanine, and proline accounted for about 80% of the total amino acids in each case. A preliminary analysis of the lipid content of this protein was made. It showed the presence of a lipid component that moves between C9 and C11 straight-chain fatty acids in the gas chromatograph. Limited amino acid sequence data were obtained by sequencing a fragment of the H.8 antigen that was isolated after partial acid hydrolysis. The H.8 antigen epitope was found to be labile to treatment with both a mild acid and a mild base.

Amino Acids↗

Monoclonal anti-galactan IgA J 539 binds intercatenarily to its polysaccharide antigen. Observations on the binding of antibody to a macromolecular antigen.

Highly purified P. Zopfii galactan of mol. wt 2 X 10(5) binds monoclonal IgA J 539 with a Ka of 5.80 X 10(5) M-1 if the polysaccharide concn is expressed in blocks of 30 galactosyl residues. This is the same Ka as found for the antibody and methyl beta (1,6)-beta-D-galactopyranosyltetraoside, the ligand capable of filling the entire combining area of the immunoglobulin. This same polysaccharide precipitates monomeric IgA J 539 on agar-double diffusion. It is concluded that the antibody binds to intercatenary galactosyl residues of the antigen.

Animals↗

Synthesis of p-nitrophenyl beta-glycosides of (1----6)-beta-D-galactopyranosyl-oligosaccharides.

Sequential tritylation, benzoylation, and detritylation of p-nitrophenyl beta-D-galactopyranoside gave p-nitrophenyl 2,3,4-tri-O-benzoyl-beta-D-galactopyranoside (2). Reaction of 2 with 2,3,4,6-tetra-O-benzoyl-alpha-D-galactopyranosyl bromide gave p-nitrophenyl O-(2,3,4,6-tetra-O-benzoyl-beta-D-galactopyranosyl)-(1----6) -2,3,4-tri-O-benzoyl-beta-D-galactopyranoside (4) in 94% yield. Deprotection with sodium methoxide then gave the crystalline p-nitrophenyl O-(beta-D-galactopyranosyl)-(1----6)-beta-D-galactopyranoside (5). Condensation of 2 with 2,3,4-tri-O-benzoyl-6-O-bromoacetyl-alpha-D-galactopyranosyl bromide (3) readily yielded the protected disaccharide p-nitrophenyl O-(2,3,4-tri-O-benzoyl-6-O-bromoacetyl-beta-D-galactopyranosyl)-(1----6) -2,3,4-tri-O-benzoyl-beta-D-galactopyranoside (6) from which the bromoacetyl groups could be selectively removed. Condensation of the resulting material with tetra-O-benzoyl-alpha-D-galactopyranosyl bromide then yielded p-nitrophenyl O-(2,3,4,6-tetra-O-benzoyl-beta-D-galactopyranosyl)-(1----6)-O-(2,3,4 -tri-O-benzoyl-beta-D-galactopyranosyl)-(1----6)-2,3,4-tri-O-benzoyl-bet a-D -galactopyranoside, (8), which was converted into the crystalline trisaccharide p-nitrophenyl O-(beta-D-galactopyranosyl)-(1----6)-O-beta-D-galactopyranosyl)-(1----6) -beta -D-galactopyranoside (9) by treatment with sodium methoxide. Preliminary experiments on the interaction of p-(bromoacetamido)phenyl and p-isothiocyanatophenyl glycoside derivatives of some of these galacto-saccharides with monoclonal anti-(1----6)-beta-D-galactopyranan antibodies have been conducted.

Antibodies, Monoclonal↗

Anticryptococcal type D antibodies raised in rabbits.

Antibodies to Cryptococcus neoformans Type D have been raised in rabbits. The partially purified antibody pool was fractionated by affinity-chromatography, and it was shown that antibodies to the capsular polysaccharide of Type D were present in the pool. Neither methyl alpha-D-mannopyranoside, nor D-mannose were inhibitors of the binding of Type D polysaccharide to the antibody-pool. Thus it appears that the serum contains antibody populations which are specific for terminal side-chain carbohydrate moieties of the immunizing antigen.

Animals↗

A surface polysaccharide of Escherichia coli O111 contains O-antigen and inhibits agglutination of cells by O-antiserum.

The repeating pentasaccharide of O-antigen from Escherichia coli O111 contains galactose, glucose, N-acetylglucosamine, and colitose, the latter representing the major antigenic determinant. Phenol extraction of this strain was previously shown to release two fractions (I and II) containing O-antigen carbohydrate, and both fractions were believed to be lipopolysaccharide. We have now characterized fractions I and II and conclude that only fraction II represents lipopolysaccharide. Fraction II contains phosphate, 2-keto-3-deoxyoctonate, beta-hydroxymyristic acid, and potent endotoxin activity, whereas fraction I was deficient in all of these properties of the lipid A and core oligosaccharide regions of lipopolysaccharide. Fractions I and II each represented 50% of the total cellular O-antigen, and both were present on the cell surface. Both fractions were metabolically stable, and no precursor-product relationship existed between them. Fraction II had a number-average molecular weight of 15,800, corresponding to an average of 12 O-antigen repeats per molecule. In contrast, fraction I had a number-average molecular weight of 354,000, corresponding to an average of 404 O-antigen repeats per molecule. Before heat treatment, cells of E. coli O111 are poorly agglutinated by O-serum; although this indicates the presence of a capsule, the corresponding K-antigen was never detected. We conclude that fraction I, when present on the cell surface, inhibits agglutination of unheated cultures of E. coli O111 by O-serum because: (i) a variant strain which lacks fraction I was agglutinated by O-serum without prior heating; (ii) erythrocytes coated with purified fraction I behaved like bacteria containing fraction I in showing inhibition of O-serum agglutination; and (iii) heat treatment released fraction I and rendered bacterial cells agglutinable in O-serum.

Agglutination↗

Structural studies on the major, capsular polysaccharide from Cryptococcus bacillisporus serotype B.

The capsular material from Cryptococcus bacillisporus serotype B has been separated into essentially two fractions. One of these (60% of the total) has been studied by the methods usual for the structural elucidation of polysaccharides. The results are consistent with a structure having nonreducing D-xylosyl as well as D-glucosyluronic acid groups attached to O-2 of D-mannosyl residues linked alpha-(1 leads to 3) in a linear backbone. Every third D-mannosyl residue is doubly substituted with a D-xylosyl group at O-2 and a D-glucosyluronic acid group at O-4.

Carbohydrate Conformation↗

The R-type lipopolysaccharides of Neisseria meningitidis.

The lipopolysaccharides of all the different serogroups of Neisseria meningitidis are of the "R" type despite the morphologically smooth appearance and the demonstrated virulence of the organisms from which they were derived. This was confirmed when each of the lipopolysaccharides was found to be devoid of detectable O-antigen side chains, giving only a low "molecular" weight core oligosaccharide when subjected to mild acid hydrolysis. The cores were modified by dephosphorylation and subjected to sugar and methylation analysis by gas-liquid chromatography. Although all the different cores contained identical components (glucose, galactose, glucosamine, heptose, and 2-keto-3-deoxyoctonate) they could be separated into three distinct categories according to their galactose:glucose ratios. These categories are typified by the cores obtained from groups A, C, and 29-e which have galactose:glucose ratios of 1:2, 2:2, and 2:1, respectively. The modified cores were methylated and analyzed by gas chromatography--mass spectrometry and on the basis of differences in the derived methylated sugars the cores could again be divided into the same three categories as above. This structural diversity also results in some serological specificity as demonstrated by the complete serogroup specificity of the group A lipopolysaccharide.

Antigens, Bacterial↗

The structure of the capsular polysaccharide from Cryptococcus neoformans serotype D.

The capsular polysaccharide from Cryptococcus neoformans serotype D has been studied by employing the usual methods for the elucidation of chemical structure. The results are consistent with the occurrence of a polysaccharide having both D-glucosyl-uronic acid and D-xylosyl groups present as nonreducing end-groups attached to O-2 of D-mannosyl residues which are linked alpha-D-(1 leads to 3) in a linear backbone.

Carbohydrate Conformation↗

Malabsorption of water miscible vitamin A in children with giardiasis and ascariasis.

Vitamin A absorption was studied using a water-miscible oral preparation of vitamin A in 19 children ages 1 1/2 to 9 years old with giardiasis and/or ascariasis, both before and after their eradication with appropriate therapy, and in three children without parasites. Marked impairment of vitamin A absorption was noted when administered in a water miscible form in children with 1) combined infection with Giardia lamblia and Ascaris lumbricoides, 2) giardiasis alone, and 3) in a proportion of children with ascariasis alone. In children with both giardiasis and ascarasis eradication of the infections promptly lead to a significant improvement in vitamin A absorption and restored it to normal. Children with giardiasis alone also showed improved vitamin A absorption after therapy. In children with ascariasis alone successful therpay did not lead to a statistically significant improvement.

Ascariasis↗

Structural elucidation of the 3-deoxy-D-manno-octulosonic acid containing meningococcal 29-e capsular polysaccharide antigen using carbon-13 nuclear magnetic resonance.

The capsular polysaccharide antigen from Neisseria meningitidis serogroup 29-e contains equimolar quantities of 2-acetamido-2-deoxy-D-galactose and 3-deoxy-D-manno-octulosonic acid (KDO), the latter of which is rarely found in biopolymers other than lipopolysaccharides. Carbon-13 nuclear mangetic resonance in conjunction with other chemical data indicated that the polysaccharide is composed of an alternating sequence of these two residues, the linkages being at C-3 of galactosamine and C-7 of KDO in the alpha-D and beta-D configuration, respectively. The native 29-e polysaccharide is O-acetylated, the O-acetyl groups being located at C-4 and C-5 of the KDO residues. Assignments of signals in the 13C nuclear magnetic resonance spectrum of the 29-e polysaccharide were made by consideration of those in the spectra of the monomer models, which necessitated the first recorded syntheses of methyl-alpha- and beta-D-3-deoxy-manno-octulopyranosonic acid. Like the methyl alpha- and beta-D-ketosides of sialic acid (Na+ salts), the equivalent methyl alpha- and beta-D-ketosides of KDO exhibit large chemical shift differences in the exocyclic C-8 position dependent on anomeric configuration. This can again be attributed to hydrogen bonding between the axial carboxylate group of the methyl beta-D anomer of KDO (C1 conformation) and the primary hydroxyl group at C-8. This phenomenon is also exhibited by the beta-D-linked KDO units of the 29-e polysaccharide.

Fourier Analysis↗