title: Novel unfolding sequence of banana lectin: Folded, unfolded and natively unfolded-like monomeric states in guanidine hydrochloride.
abstract: The sequence of unfolding events of dimeric banana lectin (Banlec), as induced by guanidine hydrochloride (GdnHCl), has been investigated by size-exclusion HPLC, fluorescence, far-UV CD, low temperature phosphorescence and selective chemical modification. 8-Anilino-1-naphthalenesulfonate (ANS) binding indicates a structured unfolding intermediate which has been characterized as dissociated monomer by size-exclusion chromatography. Interestingly, the unfolding elution pattern reveals two distinct unfolded states. One is a usual random coil. The other represents a novel species having elution behavior and structural compactness (Stokes radius) similar to dissociated monomer but showing no regular secondary structure as determined by far-UV CD, thus resembling a natively unfolded state. N-Bromosuccinimide (NBS) oxidation shows that single tryptophan residue remains unmodified in dissociated monomer intermediate while the same is oxidized in natively unfolded-like species. Such difference in tryptophan environment in these species is supported by acrylamide quenching studies, and phosphorescence results at 77 K which show a blue-shift of (0,0) band from 414.8 nm to 409.2 nm. The present results reveal subtlety of structural characteristics of unfolded states of Banlec in GdnHCl, which provide important insight in protein unfolding reaction.
authors: Department of Chemistry, Presidency University, 86/1 College Street, Kolkata 700 073, India.
authors: Goutam
authors: G
authors: Ghosh
authors: Department of Chemistry, Presidency University, 86/1 College Street, Kolkata 700 073, India. Electronic address: dm.pcchem@gmail.com.
authors: Dipak K
authors: DK
authors: Mandal
mesh_terms: D000595:Amino Acid Sequence; D002942:Circular Dichroism; D019791:Guanidine / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D028521:Musa / Q000737:chemistry*; D037121:Plant Lectins / Q000737:chemistry*; D011489:Protein Denaturation; D017433:Protein Structure, Secondary
chemical_list: D037121:Plant Lectins; D019791:Guanidine
keywords: Banana lectin; Natively unfolded state; Random coil; Structured intermediate; Unfolding sequence