Publications (6)15.5 Total impact
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Article: Stereo-selectivity of human serum albumin to enantiomeric and isoelectronic pollutants dissected by spectroscopy, calorimetry and bioinformatics.
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ABSTRACT: 1-naphthol (1N), 2-naphthol (2N) and 8-quinolinol (8H) are general water pollutants. 1N and 2N are the configurational enantiomers and 8H is isoelectronic to 1N and 2N. These pollutants when ingested are transported in the blood by proteins like human serum albumin (HSA). Binding of these pollutants to HSA has been explored to elucidate the specific selectivity of molecular recognition by this multiligand binding protein. The association constants (K(b)) of these pollutants to HSA were moderate (10(4)-10(5) M(-1)). The proximity of the ligands to HSA is also revealed by their average binding distance, r, which is estimated to be in the range of 4.39-5.37 nm. The binding free energy (ΔG) in each case remains effectively the same for each site because of enthalpy-entropy compensation (EEC). The difference observed between ΔC(p) (exp) and ΔC(p) (calc) are suggested to be caused by binding-induced flexibility changes in the HSA. Efforts are also made to elaborate the differences observed in binding isotherms obtained through multiple approaches of calorimetry, spectroscopy and bioinformatics. We suggest that difference in dissociation constants of pollutants by calorimetry, spectroscopic and computational approaches could correspond to occurrence of different set of populations of pollutants having different molecular characteristics in ground state and excited state. Furthermore, our observation of enhanced binding of pollutants (2N and 8H) in the presence of hemin signifies that ligands like hemin may enhance the storage period of these pollutants in blood that may even facilitate the ill effects of these pollutants.PLoS ONE 01/2011; 6(11):e26186. · 4.09 Impact Factor -
Article: Purification and characterization of mannose/glucose-specific lectin from seeds of Trigonella foenumgraecum.
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ABSTRACT: A lectin present in seeds of Trigonella foenumgraecum was isolated and purified by acid precipitation, salt fractionation, and affinity chromatography on mannan cross-linked agarose. SDS-PAGE revealed a single band corresponding to a molecular weight of 27,350 daltons. The lectin agglutinated trypsin-treated rat erythrocytes. Sugar specificity as determined by hemagglutination inhibition assay indicated that the lectin belongs to a glucose/mannose-specific group. The reaction of the lectin with glycoprotein was affected by pH changes. The carbohydrate binding specificity of the lectin was investigated by turbidity and activity measurements. As the lectin belongs to the Leguminoceae family, the specificity of the lectin for glucose/mannose renders it a valuable tool for Rhizobium-legume symbiosis.Biochemistry (Moscow) 02/2007; 72(1):44-8. · 1.06 Impact Factor -
Article: Comparative study of effects of polyols, salts, and alcohols on trichloroacetic acid-induced state of cytochrome C.
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ABSTRACT: A systematic investigation of the effect of polyethylene glycols, salts, and alcohols on the trichloroacetic acid (TCA)-induced state of ferricytochrome c was made using various spectroscopic techniques. Native cytochrome c (Cyt c) has a fluorescence maximum at 335 nm, whereas the TCA-induced state of Cyt c has a red shift of 7 nm with enhanced fluorescence intensity. The near- and far-UV CD spectra showed a significant loss of tertiary and secondary structure, although the protein is relatively less unfolded as compared with a conformation at pH 2.0. Addition of 70% (v/v) polyols to TCA (3.3 mM)-induced state of Cyt c resulted in increased 1-anilino-8-naphthalene sulfonate binding and increased mean residue ellipticity at 222 nm, indicating increase in compactness with enhanced exposure of hydrophobic surface area. Also, the stabilizing effect of salts and alcohols on the TCA-induced state was studied and compared with their effect on trifluoroacetic acid-unfolded state of Cyt c. Among all the polyols, salts, and alcohols studied, PEG-400, K3[Fe(CN)6], and butanol were the most efficient in inducing secondary structure in TCA-induced state as examined by the above-mentioned spectroscopic techniques. For salts, the efficiency in inducing the secondary structure followed the order K3[Fe(CN)6] > KClO4 > K2SO4 > KCl. For alcohols, this order was found to be as follows: butanol > propanol > ethanol > methanol.Biochemistry (Moscow) 11/2006; 71(10):1101-9. · 1.06 Impact Factor -
Article: A tuber lectin from Arisaema helleborifolium Schott with anti-insect activity against melon fruit fly, Bactrocera cucurbitae (Coquillett) and anti-cancer effect on human cancer cell lines.
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ABSTRACT: An anti-insect and anti-cancer lectin has been isolated from Arisaema helleborifolium Schott by affinity chromatography using asialofetuin-linked amino activated silica beads. The bound A. helleborifolium lectin (AHL) was eluted with 100mM glycine-HCl buffer, pH 2.5. It gave a single band on SDS-PAGE, pH 8.3, and PAGE, pH 4.5. However, multiple bands were obtained in PAGE at pH 8.3 and isoelectric focusing. The lectin was a homotetramer having subunit molecular mass 13.4kDa while its native molecular mass was 52kDa. It was a glycoprotein with 3.40% carbohydrate and was stable up to 60 degrees C for 30min. It showed anti-insect activity towards second instar larvae of Bactrocera cucurbitae (Coquillett) with LC(50) value of 16.4microg/ml. Larvae fed on artificial diet containing sub-lethal dose of AHL showed a significant decrease in acid phosphatase and alkaline phosphatase activity while esterase activity markedly increased as compared to larvae fed on diet without lectin. AHL was also found to inhibit in vitro proliferation of some well established human cancer cell lines viz HOP-62 (95%), HCT-15 (92%), HEP-2 (66%), HT-29 (68%), PC-3 (39.4%), and A-549 (20.7%).Archives of Biochemistry and Biophysics 02/2006; 445(1):156-65. · 2.93 Impact Factor -
Article: Molten globule-like folding intermediate of asialofetuin at acidic pH.
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ABSTRACT: In our earlier communication on acid-induced unfolding of bovine serum fetuin (BSF), we showed the existence of a molten globule (MG)-like state of BSF at pH 1.8. The MG state was characterized by higher content of secondary structure than native and almost complete loss of tertiary structure and more solvent exposed hydrophobic surface [Biochim. Biophys. Acta 1649 (2003) 164]. In this work we have shown the presence of an MG-like partially folded intermediate of asialofetuin at around pH 1.8, which is much different from the MG state observed in BSF in secondary structure contents. The results show that asialofetuin at pH 1.8 retains approximately 45% secondary structure, as evident from far-UV CD spectra. The near-UV CD spectra showed almost complete loss of tertiary structure. The intrinsic fluorescence and acrylamide quenching of the lone tryptophan residue showed that in acid-induced state, it is buried in the interior in a nonpolar environment. The temperature dependence of far-UV CD signal of asialofetuin at pH 1.8 exhibits a weak cooperative thermal transition. A significant increase in ANS fluorescence showed extensive solvent exposure of nonpolar cluster. Size exclusion chromatography (SEC) indicates a slight increase in the hydrodynamic size of acid-induced protein. These results suggest that asialofetuin at pH 1.8 represents the MG-like folding intermediate. Moreover, our results showed that glycosylation might play a role in stabilization of secondary structure during acid and/or thermal denaturation.Biochimica et Biophysica Acta 07/2004; 1699(1-2):191-9. · 4.66 Impact Factor -
Article: Mitogenic lectins.
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ABSTRACT: Lectins have captured the attention of a large number of researchers on account of the various exploitable activities that they exhibit, including their proliferative effects on various cell types. Recognition of cell-surface carbohydrates by lectins has broad implications in important biological processes. The ability of lectins to detect subtle variations in carbohydrate structures on the surface of cells and tissues has made them a paradigm for protein-carbohydrate recognition. Lectins display a considerable repertoire of carbohydrate specificities. These characteristics, together with the ability to stimulate lymphocytes as well as other cells, have made lectins an important diagnostic and experimental tool to study the various aspects of cell growth and differentiation, taking lymphocytes as the cell type. Various models of lectin action have been used to explain specific event-driven functions of lectins, which have convincingly elucidated the mechanism of action of lectins in different milieu. This review contains a selective survey of information available on the role of lectins in cell proliferation, with special emphasis on lymphocyte proliferation and the models which have been proposed to elucidate lectin action. The information available clearly justifies the observation that a generalized scheme to explain the diverse roles played by lectins in cell growth and differentiation is still awaited.Medical science monitor: international medical journal of experimental and clinical research 12/2003; 9(11):RA265-9. · 1.70 Impact Factor
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Institutions
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2003–2011
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Aligarh Muslim University
- Interdisciplinary Biotechnology Unit
Alīgarh, Uttar Pradesh, India
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