01792nas a2200265   4500008004100000022001400041245008800055210006900143260007300212300001200285490000700297520083600304653001601140653003501156653001701191653001901208653004801227653001401275100002801289700002901317700002301346700002201369700002501391856011001416       2013                        eng d  a0141-813000aSolution and in silico studies on the recombinant lectin from Cicer arietinum seeds0 aSolution and in silico studies on the recombinant lectin from Ci  aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcMAY  a149-1550 v563 a<p>The Cicer arietinum seed lectin was cloned and expressed in Escherichia coli and purified in active form. Conformational characterization of the recombinant lectin (rCAL) was performed using biophysical and bioinformatics tools. Thermal denaturation of rCAL caused rapid secondary structural rearrangements above 50 degrees C and transient exposure of hydrophobic residues at 55 degrees C, leading to aggregation. Treatment of rCAL with GdnHCl resulted in unfolding followed by dissociation of the dimer. The single tryptophan in rCAL present on the surface of the protein is surrounded by hydrophobic and acidic amino acids and exists as different conformers. The experimental observations correlated well with the structural information revealed from the homology model of rCAL. (C) 2013 Elsevier B.V. All rights reserved.</p>10aAggregation10aBL21-CodonPlus (DE3)-RIL cells10aCicer lectin10aHomology model10aSteady-state and time resolved fluorescence10aUnfolding1 aWakankar, Madhurima, S.1 aKrishnasastry, Musti, V.1 aJaokar, Tulika, M.1 aPatel, Krunal, A.1 aGaikwad, Sushama, M.  uhttps://library.ncl.res.in/content/solution-and-silico-studies-recombinant-lectin-cicer-arietinum-seeds-002449nas a2200265   4500008004100000022001400041245010100055210006900156260000800225300001000233490000700243520157500250653001701825653001201842653001901854653001901873653003301892653002401925100001701949700002001966700002601986700002002012700002102032856013002053       2016                        eng d  a0141-813000aStructural-functional insights and studies on saccharide binding of Sophora japonica seed lectin0 aStructuralfunctional insights and studies on saccharide binding   cOCT  a75-840 v913 a<p>Functional and conformational transitions of the Sophora japonica seed lectin (SJL) were studied in detail using bioinformatics and biophysical tools. Homology model of the lectin displayed all the characteristics of the legume lectin monomer and the experimental observations correlated well with the structural information. In silico studies were performed by protein-ligand docking, calculating the respective binding energies and the residues involved in the interactions were derived from LigPlot(+) analysis. Fluorescence titrations showed three times higher affinity of T-antigen disaccharide than N-acetyl galactosamine (GaINAc) towards SJL indicating extended sugar binding site of the lectin. Thermodynamic parameters of T-antigen binding to SJL indicated the process to be endothermic and entropically driven while those of GaINAc showed biphasic process. SDS-PAGE showed post-translationally modified homotetrameric species of the lectin under native conditions. In presence of guanidine hydrochloride (0.5-5.0 M), the tetramer first dissociated into dimers followed by unfolding of the protein as indicated by size exclusion chromatography, fluorescence and CD spectroscopy. Different structural rearrangements were observed during thermal denaturation of SJL at physiological pH 7.2, native pH 8.5 and molten globule inducing pH 1.0. Topological information revealed by solute quenching studies at respective pH indicated differential hydrophobic environment and charge density around tryptophan residues. (C) 2016 Elsevier B.V. All rights reserved.</p>10aDissociation10adocking10aHomology model10aLigand binding10aSophora japonica seed lectin10aThermal transitions1 aYadav, Priya1 aShahane, Ganesh1 aRamasamy, Sureshkumar1 aSengupta, Durba1 aGaikwad, Sushama  uhttps://library.ncl.res.in/content/structural-functional-insights-and-studies-saccharide-binding-sophora-japonica-seed-lectin02220nas a2200241   4500008004100000022001400041245012000055210006900175260000900244300001200253490000800265520138800273653001701661653001901678653001101697653001901708653002301727653002401750100002801774700002101802700002501823856013001848       2019                        eng d  a0003-986100aInvestigation of structural and saccharide binding transitions of Bauhinia purpurea and Wisteria floribunda lectins0 aInvestigation of structural and saccharide binding transitions o  cFEB   a134-1420 v6623 a<p>Two novel medicinally important legume lectins from Bauhinia purpurea (BPL) and Wisteria floribunda (WFL) possessing extended sugar binding site were investigated for functional and conformational transitions using biochemical and biophysical techniques as well as bioinformatical tools. Homology model of BPL was constructed using the Schrodinger suite and docked with N-acetyl galactosamine and T-antigen disaccharide (Gal beta 1-3GalNAc alpha O-Me). The longer loop D in the structure of WFL compared to that in BPL was found to be responsible for its specificity to LacdiNac (beta-D-GalNAc-[1 -&gt; 4]-DGlcNAc) over Gal beta 1-3GalNAc. BPL remained functionally stable up to 40 degrees C whereas WFL remained stable upto 70 degrees C indicating the strength of the sugar binding site geometry. Both the lectins showed intense but non-specific secondary structure in the range of 65-90 degrees C. WFL showed rapid aggregation above 80 degrees C as indicated by light scattering intensity. The lectins showed simultaneous dissociation and multistate unfolding in the vicinity of GdnHCl. At pH 1.0, both the lectins exhibited molten globule like structures, which were characterized further and were found to respond in a different way towards denaturants. The results have provided valuable insights into the molecular basis of the activity and stability of the two lectins.</p>10aConformation10aHomology model10aLectin10aMolten globule10asaccharide binding10aThermal aggregation1 aAgrawal, Sanskruthi, B.1 aGhosh, Deepanjan1 aGaikwad, Sushama, M.  uhttps://library.ncl.res.in/content/investigation-structural-and-saccharide-binding-transitions-bauhinia-purpurea-and-wisteria