title: Solution and in silico studies on the recombinant lectin from Cicer arietinum seeds.
abstract: 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 °C and transient exposure of hydrophobic residues at 55 °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.
authors.affiliation: Division of Biochemical Sciences, National Chemical Laboratory, Pune, Maharashtra, 411 008, India.
authors.forename: Madhurima S
authors.initials: MS
authors.lastname: Wakankar
authors.forename: Musti V
authors.initials: MV
authors.lastname: Krishnasastry
authors.forename: Tulika M
authors.initials: TM
authors.lastname: Jaokar
authors.forename: Krunal A
authors.initials: KA
authors.lastname: Patel
authors.forename: Sushama M
authors.initials: SM
authors.lastname: Gaikwad
mesh_terms: D000818:Animals; D029863:Cicer / Q000737:chemistry* / Q000187:drug effects; D003001:Cloning, Molecular; D003198:Computer Simulation; D019791:Guanidine / Q000494:pharmacology; D037102:Lectins / Q000737:chemistry* / Q000302:isolation & purification; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011489:Protein Denaturation / Q000187:drug effects; D058849:Protein Refolding / Q000187:drug effects; D017433:Protein Structure, Secondary; D058767:Protein Unfolding / Q000187:drug effects; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D015203:Reproducibility of Results; D012639:Seeds / Q000737:chemistry*; D012996:Solutions; D013050:Spectrometry, Fluorescence; D040681:Structural Homology, Protein; D013696:Temperature; D014364:Tryptophan / Q000378:metabolism
chemical_list: D037102:Lectins; D011994:Recombinant Proteins; D012996:Solutions; D014364:Tryptophan; D019791:Guanidine