Biomedical subjects

B P Chatterjee

Publications and source records attributed to B P Chatterjee.

At least 19 recordsLinked to original sources

PubMed1995

Saracin: a lectin from Saraca indica seed integument recognizes complex carbohydrates.

A lectin isolated from Saraca indica seed integument was purified by affinity chromatography on porcine thyroglobulin Sepharose followed by Sephadex G-50 and shown to be homogeneous by PAGE. It showed a single band on SDS-PAGE in the absence and presence of 2-mercaptoethanol corresponding to a M(r) of congruent to 12,000, thus indicating it to be a monomer. The lectin agglutinated erythrocytes of human A, B, O and AB blood group, animal erythrocytes as well as Ehrlich ascites cells. It is a thermostable glycoprotein containing 11.6% carbohydrates and large proportions of acidic amino acids. In haemagglutination-inhibition assays, among the tested glycoproteins, porcine thyroglobulin having the NeuAc alpha (2-6)/(2-3)D-Gal beta (1-4)D-GlcNAc sequence was found to be the most potent; however, its asialo counterpart was non-inhibitory. The lectin is present solely in the seed integument even in the immature stage. During maturation of the seed the lectin activity declined and was completely absent when totally matured and dried. Studies in vitro showed that on incubation at 37 degrees the seed gradually lost its lectin activity which was completely absent after 62 days with 88.5% loss of water. Similar studies on scraped seed integument revealed that the lectin activity was lost in 22 days with 87.5% dehydration.

ABO Blood-Group System↗

PubMed1995

Cocos nucifera pollen inducing allergy: sensitivity test and immunological study.

Among the heterogeneous population (n = 975) in greater Calutta, sensitization to Cocos nucifera pollen accounts to be 2.65% and for atopic patients (n = 204) 47.06%. Out of 24 patients who had C. nucifera pollen sensitivity and suffered from asthma and allergic rhinitis, 16 showed sensitivity also to other allergens. All were skin test positive and 19 of them were phadezym RAST positive to C. nucifera pollen extract. Bronchial provocation test appeared to be positive in 7 out of 8 patients included in the test and no late response or non-specific reactions were observed. C. nucifera pollen extract on fractionation by ion-exchange chromatography following gel filtration yielded two major allergenic protein fractions, CnII (M(r) 158,000) and CnVII (M(r) 2900) as evidenced by skin prick test, ELISA-inhibition and immunoblot analysis. Hence, C. nucifera pollen should be considered to be a relevant allergen and thus included in the panel of allergens for routine clinical use.

Adolescent↗

PubMed1994

Isolation and characterization of two IgE-reactive proteins from Azadirachta indica pollen.

Two allergenically active components present in the Azadirachta indica whole pollen extract have been isolated by sequential ammonium sulfate precipitation (0-90%), DEAE-Sephadex A-50 ion-exchange chromatography followed by gel filtration through Sephadex G-200. The allergenicity of fractionated materials has been tested by skin prick test and ELISA inhibition which reveal that AIaI and AIaIVb are the major allergens. Immunoblot confirms the IgE-binding activity of the proteins. Although both fractions are found to be homogeneous by SDS-PAGE, isoelectric focusing produces more than one isoelectric point in AIaI (pI = 3.15, 3.3 and 3.5) and AIaIVb (pI = 6.0 and 6.2). Amino acid analyses of the two allergens, the effect of pH on them and cross-reactivity between them have been discussed.

Allergens↗

PubMed1994

Typing of Shigella dysenteriae strains of different serogroups by lectins.

Ten different serogroups of Shigella dysenteriae were typed with the aid of lectins of known sugar specificity resulting from their interactions with the carbohydrates on lipopolysaccharides in the outer membrane of bacteria as evidenced by the agglutination-inhibition assay with simple carbohydrates. Lipopolysaccharides of two serogroups of Shigella were precipitated with different lectins and the results were corroborated by those derived from the agglutination assay suggesting that Shigella dysenteriae can be characterized on the species level with the aid of lectins.

Agglutination↗

PubMed1993

Purification, characterisation, and carbohydrate specificity of the lectin of Ficus cunia.

A lectin, isolated from the seeds of Ficus cunia and purified by affinity chromatography on fetuin-Sepharose, was homogeneous in PAGE, GPC, HPLC, and immunodiffusion, and had mol wt of 3200-3500. In SDS-PAGE and HPLC in the absence and presence of 2-mercaptoethanol, the lectin gave a single band or peak corresponding to M(r) 3300-3500, thus indicating it to be a monomer. The lectin agglutinated human erythrocytes regardless of blood group, bound to Ehrlich ascites cells and to human rat spermatozoa, and was thermally stable; its activity was enhanced by Ca2+. The lectin is a metalloprotein that was inactivated by dialysis with EDTA followed by acetic acid, but reactivated by the addition of Ca2+. The lectin contained 2.0% of carbohydrates, large proportions of acidic amino acids, but little methionine. In hapten-inhibition assays, chitin oligosaccharides [(1-->4)-linked beta-GlcNAc] and N-acetyl-lactosamine were inhibitors of which N,N',N",N"'-tetra-acetylchitotetraose was the most potent. Among the macromolecules tested that contain either multiple N-acetyl-lactosamine and/or (1-->4)/(1-->6)-linked beta-GlcNAc, asialofetuin glycopeptide was the most potent inhibitor. Thus, an N-acetyl group and substitution at C-1 of D-GlcN are necessary for binding.

Animals↗

PubMed1993

Artocarpin-galactomannan interaction: characterization of combining site of artocarpin.

Artocarpin, a lectin purified from Artocarpus lakoocha seeds, precipitates well several galactomannans having terminal D-Gal-alpha-(1-6)-residues. Quantitative precipitin-inhibition studies using various haptens suggest that the -OCH2-group at C-1 and hydroxyl groups at C-4, and partially at C-6, in the alpha-glycoside of D-galactose configuration are necessary for lectin-sugar interaction. The formation of an artocarpin-fenugreek galactomannan complex was dependent on temperature, pH, ionic strength and the number of hydroxy groups of alditols added to the medium.

Binding Sites↗

PubMed1992

Effects of chemical reagents on the allergenicity of house dust.

Con A-Sepharose bound and unbound house dust allergens were treated with various protein modifying reagents in order to ascertain the amino acid residues responsible for their allergenicity. Modification of carboxy group and arginine residue of bound fraction led to 90 and 85% loss of allergenicity whereas carboxy group, lysine and tryptophan modified unbound allergen reduced greater than or equal to 85% of activity. Allergenicity of both fractions was highly affected by pronase. On periodate treatment the bound fraction lost its allergenicity more compared to unbound one. Chemical regents had no remarkable effect on the activity of unbound fraction.

Allergens↗

PubMed1991

Chemical modification studies of Artocarpus lakoocha lectin artocarpin.

The effect of chemical modification on an anti T-like lectin, artocarpin isolated from Artocarpus lakoocha seeds was investigated in order to identify the type of amino acids involved in its agglutinating activity. Modification of carboxyl groups, arginine and lysine residues, did not affect the lectin activity. However, modification of tryptophan, tyrosine and histidine residues led to a complete loss of its activity, indicating the involvement of these amino acids in the saccharide-binding ability. A protection was observed in the presence of inhibitory sugar. A marked decrease in the fluorescence emission was found when the tryptophan residues of lectin were modified. The circular dichroism spectra showed the presence of an identical pattern of conformation in the native and modified lectin, indicating that the loss in activity was due to modification only. The effect of pronase on artocarpin showed loss of activity whereas papain and trypsin had no effect. The specific activity of artocarpin remained unaltered on treatment with glycosidases but remarkable increase in the activity (of the same) was observed with xylanase treatment. Immunodiffusion studies with chemically modified lectin showed no gross structural changes, indicating that the group specific modifying agents did not alter the antigenic sites of the modified lectin.

Amino Acids↗

PubMed1991

Studies on the effect of various parameters on crotalarin-fetuin interaction.

With a view to optimise the interaction of crotalarin, a blood group A-specific lectin from the seeds of Crotalaria striata with fetuin the effect of various parameters on the reaction has been studied turbidimetrically. The formation of crotalarin-fetuin complex was dependent on time, temperature, pH and the ionic strength of the medium. The maximum turbidity appeared in 30 min at 20 degrees C and the pH optimum was 3.5. The binding constant (Ka) for crotalarin-fetuin interaction was 5.58 x 10(4) M-1 (pH 3.5) at 20 degrees C. Among the different inorganic salts tested, the cations with increasing concentrations had pronounced effect on binding. KCNS and KI, however, were noninhibitory. The turbidity slightly increased in presence of different sodium salts, whereas periodate and urea reduced the interaction. The different alcohols had no remarkable effect on the above reaction.

Hydrogen-Ion Concentration↗

PubMed1990

Chemical modification studies on a blood group A-specific lectin, crotalarin (Crotalaria striata) and its effect on hemagglutinating activity.

Crotalarin, the N-acetyl-D-galactosamine-binding blood group A-specific lectin from the seeds of Crotalaria striata was subjected to various chemical modifications in order to ascertain the amino acid residues responsible for its carbohydrate-binding property. Modification of lysine, cysteine and arginine residues did not affect the carbohydrate-binding activity of the lectin. However, modification of tyrosine residue and carboxy group of the acidic amino acids led to a complete loss of its activity, indicating the involvement of tyrosine and aspartic and glutamic acid in the saccharide-binding respectively. The hemagglutinating activity of the lectin was completely/almost completely lost by modification of tryptophan residues. The relative loss in hemagglutinating activity on modification of tryptophan residues indicate that one residue/molecule is required for the carbohydrate-binding activity of the lectin. Modification was not effective in the presence of D-galactose (0.2 M). A marked decrease in the fluorescence emission was found as the tryptophan residues of crotalarin were modified. The c.d. spectra showed the presence of an identical pattern of conformation in the native and modified lectins which confirms that the loss in activity was due to modification only. The effect of periodate oxidation on crotalarin showed loss of activity whereas action of enzymes retained most of the activity.

ABO Blood-Group System↗

PubMed1990

Isolation of concanavalin A-reactive low-molecular weight allergen from house dust.

A concanavalin A-reactive low-molecular weight (MW approximately 7000) glycoprotein allergen has been isolated from house dust. The crude allergen was fractionated by Sephadex gel filtration followed by separation into two allergenic components by affinity chromatography on concanavalin A-Sepharose column. The column-bound fraction contains Man (11.7%), Gal (2.34%), GlcNAc (3.47%) and a trace of Glc and has been proved to be more potent allergen for house dust-sensitive patients both in vivo and in vitro tests.

Allergens↗

PubMed1989

Further characterization and immunochemical studies on the carbohydrate specificity of jackfruit (Artocarpus integrifolia) lectin.

The lectin from jackfruit (Artocarpus integrifolia) seeds has been purified by Rivanol (6,9-diamino-2-ethoxyacridine lactate) treatment. The specific activity, molecular weights of parent lectin and its subunit, its glycoprotein nature, and hemagglutination-inhibition assays suggest that this preparation is identical to that obtained by affinity chromatography on melibiose-agarose adsorbent (Ahmed, H., and Chatterjee, B. P. (1986) in Lectins, Biology, Biochemistry, Clinical Biochemistry (Bøg-Hansen, T. C., and van Driessche, E., eds) Vol. 5, pp. 125-133, Walter de Gruyter, New York). The lectin strongly agglutinates human and several animal erythrocytes. The lectin contains five isolectins of pI values 7.1, 6.85, 5.5, 5.3, and 5.1. It is thermally stable and loses its activity above 75 degrees C. The hemagglutinating activity remains unchanged in the presence of bivalent cations viz., Ca2+, Mg2+, Mn2+, etc. It is a metalloprotein. The lectin retains its activity by dialysis with acetic acid followed by EDTA. It agglutinates Ehrlich ascites cells. Equilibrium dialysis of lectin with melibiose and quenching of fluorescence of 4-methylumbelliferyl-alpha-D-galactopyranoside by the lectin show that homotetrameric jackfruit lectin has two sugar-binding sites. The lectin precipitates well several galactomannans and glycoproteins having terminal D-Gal-alpha-(1----6)- or D-Gal-beta-(1----3)-D-GalNAc residues. It hardly or does not precipitate polysaccharides having terminal D-Gal-alpha-(1----3) residues. Quantitative precipitin-inhibition studies using various haptens suggest that the -OCH2- group at C-1 and -OH groups at C-4 and partially at C-6 in the alpha-glycoside of D-galactose configuration are important for lectin-sugar interaction.

Animals↗

PubMed1989

Lectin typing of Pseudomonas aeruginosa strains of different serogroups, Habs and Fisher types.

Sixteen Habs and three Fisher types of Pseudomonas aeruginosa were typed with lectins of know specificity resulting from their interaction with bacterial cell surface carbohydrates as evidenced by agglutination-inhibition assay with simple carbohydrates. Lipopolysaccharides of few strains of Pseudomonas are precipitated with different lectins and the results are corroborated by those of agglutination suggesting that Pseudomonas aeruginosa can be characterized intraspecifically by lectins.

Agglutination Tests↗

PubMed1988

Wistaria sinensis agglutinin: purification, carbohydrate specificity, and characterisation of the combining site.

A 2-acetamido-2-deoxy-D-galactose-binding agglutinin from Wistaria sinensis seeds, purified by affinity chromatography on a 2-acetamido-2-deoxy-D-galactose-starch conjugate, was homogeneous as judged by poly(acrylamide) disc gel electrophoresis. It had a mol. wt. of 66,000 (gel filtration on Sephadex G-150); on electrophoresis on SDS-poly(acrylamide) gel in the presence of 2-mercaptoethanol, it dissociated into sub-units of mol. wt. 34,000, suggesting the agglutinin to be a dimer; and it was a glycoprotein containing 4.8% of carbohydrate. It agglutinated several vertebrate erythrocytes, including human regardless of the blood group. In hapten-inhibition assays, 2-acetamido-2-deoxy-D-galactose and its glycosides were found to be better inhibitors than D-galactose and its glycosides, but N-acetyl-lactosamine was the most potent inhibitor. The binding involved HO-3,4 of the haptens and HO-2 partially.

Animals↗

PubMed1987

Hemagglutination patterns of Pseudomonas aeruginosa strains of different serogroups, Habs and Fisher types.

Pseudomonas aeruginosa strains of sixteen Habs and three Fisher types were examined for their hemagglutination activity using vertebrate erythrocytes including human ones. Their carbohydrate-binding specificity as determined by hemagglutination-inhibition suggests a possible method of their purification. This study also indicates the possibility of an occurrence of multiple lectins of different specificity.

Hemagglutination Inhibition Tests↗

PubMed1986

Serological characterization of humoral lectin from Heterometrus granulomanus scorpion hemolymph.

A lectin in the hemolymph of Indian scorpion Heterometrus granulomanus was detected by agglutination of human and animal erythrocytes. The agglutinating activity was enhanced in presence of Ca2+ ion. The lectin shows specificity for sialic acid like many other sialic acid-specific lectins such as "Limulin", as the agglutination of erythrocytes was completely abolished by treatment with Vibrio cholerae neuraminidase. However, neuraminidase-treated rat and mouse erythrocytes exhibited high titers. This result was substantiated by crossed-absorption test which suggests the adjunct specificity of the Heterometrus lectin for these cells. Hemagglutination-inhibition results of the lectin indicate that sialic acid including its derivatives and sialoglycoconjugates are the inhibitors among which glycophorin was most potent.

Animals↗

PubMed1986

Purification of a sialic acid-specific lectin from the Indian scorpion Heterometrus granulomanus.

A sialic acid-specific lectin, scorpin, has been purified to apparent homogeneity from the Indian scorpion Heterometrus granulomanus by affinity chromatography on equine submandibular gland glycopeptides linked to Sepharose and gel filtration on Sephadex G-200. The lectin has a molecular mass of 500 000 Da and was dissociated into single polypeptide chains of 15 000 Da, as determined by SDS gel electrophoresis in the presence of 2-mercaptoethanol. Scorpin is a glycoprotein containing 2.8% sugars. Its specificity was investigated by the inhibition of hemagglutination with various derivatives of sialic acid and other sugars. N-Acetylneuraminic acid gave better inhibition than N-glycoloylneuraminic acid but showed less inhibitory effect than sialyl-alpha(2----3)-lactose and disialyllactose. Among the sialoglycoconjugates tested, equine submandibular gland glycopeptide was found to be the most potent inhibitor. Scorpin showed a strong tendency to bind to carboxyl groups, since reduction of the carboxyl group of N-acetylneuraminic acid destroyed the inhibitory potency of this sugar. Furthermore, D-glucuronic acid inhibited hemagglutination whereas N-acetylglucosamine or N-acetylgalactosamine were not inhibitors.

Animals↗

PubMed1985

Jackfruit (Artocarpus integrifolia) and the Agaricus mushroom lectin fit also to the so-called peanut receptor.

The lectin from jackfruit (Artocarpus integrifolia) reacts with the peanut-specific lectin receptor represented by the T-disaccharide, which is also part of the Thomsen (= T)-Friedenreich antigen, namely O-beta-D-galactosyl-(1----3)-N-acetyl-D-galactosamine. The difference between the jackfruit lectin and peanut lectin has been shown to be mainly due to the further linkage of the T-disaccharide: Whereas the peanut lectin reacts with both anomeric forms, jackfruit lectin and the closely - with respect to its specificity - related Agaricus bisporus mushroom lectin react predominantly with the alpha-linked T-disaccharide, so that they cannot detect the lipid-linked T-disaccharide, as the peanut lectin does.

Antigens, Tumor-Associated, Carbohydrate↗