title: A new chitooligosaccharide specific lectin from snake gourd (Trichosanthes anguina) phloem exudate. Purification, physico-chemical characterization and thermodynamics of saccharide binding.
abstract: A new lectin has been purified to homogeneity from the phloem exudate of snake gourd (Trichosanthes anguina) by affinity chromatography on chitin. The snake gourd phloem lectin (SGPL) specifically binds chitooligosaccharides and their inhibitory potency increased with increase in size. PAGE and SDS-PAGE studies indicate that SGPL is a heterodimer, in which the two subunits (48 and 53kDa) are joined by disulfide bonds. Consistent with this, electrospray-ionization mass spectrum yielded the exact mass of the protein as 104,621.8 Daltons. CD studies showed that SGPL contains about 9% α-helix, 39% β-sheet, 20% β-turns and 32% unordered structures and that saccharide binding does not significantly affect its secondary and tertiary structures. Titration calorimetric studies indicate that the dimeric lectin binds two ligand molecules [(GlcNAc)(3-6)] with association constants determined at 25°C being 1.7×10(5) and 3.6×10(5)M(-1), for chitotriose and chitohexaose, respectively. Binding of all the chitooligosaccharides is governed by enthalpic forces, whereas the contribution from binding entropies was unfavorable. These results suggest that the SGPL-saccharide interaction is stabilized by hydrogen bonding and van der Waals' interactions. Enthalpy-entropy compensation was observed for the SGPL-chitooligosaccharide interaction, suggesting that water molecules play a key role in the binding process.
authors.affiliation: School of Chemistry, University of Hyderabad, Central University P.O., Gachi Bowli, Hyderabad, Andhra Pradesh 500046, India.
authors.forename: Akkaladevi
authors.initials: A
authors.lastname: Narahari
authors.forename: Pavan Kumar
authors.initials: PK
authors.lastname: Nareddy
authors.forename: Musti J
authors.initials: MJ
authors.lastname: Swamy
mesh_terms: D000818:Animals; D002152:Calorimetry, Differential Scanning; D002241:Carbohydrates; D002846:Chromatography, Affinity; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism*; D037121:Plant Lectins / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D013816:Thermodynamics; D029778:Trichosanthes / Q000737:chemistry*
chemical_list: D002241:Carbohydrates; D009844:Oligosaccharides; D037121:Plant Lectins; C091028:phloem lectin