title: Differing structural characteristics of molten globule intermediate of peanut lectin in urea and guanidine-HCl.
abstract: The structural characteristics of exclusive equilibrium molten globule-like intermediate formed during peanut lectin unfolding in urea and guanidine hydrochloride (GdnHCl) have been investigated by size-exclusion chromatography, circular dichroism, fluorescence, phosphorescence, and chemical modification. The elution behavior and 8-anilino-1-naphthalenesulfonate binding indicate a less compact tertiary structure in urea than in GdnHCl. Further, the urea-induced intermediate reveals perturbed, nonnative typical β-sheet conformation in contrast to native-like atypical β-structure in GdnHCl. N-bromosuccinimide oxidation shows that none of three tryptophan residues is modified for GdnHCl-induced intermediate while one gets oxidized in urea. Such difference in tryptophan environment is supported by acrylamide quenching (Stern-Volmer constant being 3.2 and 5.8 M(-1) respectively), and phosphorescence studies at 77 K which show a blue-shift of (0, 0) band from 412.4 nm (GdnHCl) to 411.4 nm (urea). These results may provide important insight into the differential effects of GdnHCl and urea on the structural characteristics of intermediate state(s) in protein folding.
authors.affiliation: Department of Chemistry & Biochemistry, Presidency University, 86/1 College Street, Kolkata 700 073, India.
authors.forename: Goutam
authors.initials: G
authors.lastname: Ghosh
authors.forename: Dipak K
authors.initials: DK
authors.lastname: Mandal
mesh_terms: D020106:Acrylamide / Q000737:chemistry; D000817:Anilino Naphthalenesulfonates / Q000737:chemistry; D001974:Bromosuccinimide / Q000737:chemistry; D019791:Guanidine / Q000737:chemistry*; D019887:Peanut Agglutinin / Q000737:chemistry*; D011485:Protein Binding; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D058767:Protein Unfolding; D013696:Temperature; D014364:Tryptophan / Q000737:chemistry; D014508:Urea / Q000737:chemistry*
chemical_list: C515594:8-anilino-1-naphthalenesulfonic acid; D000817:Anilino Naphthalenesulfonates; D019887:Peanut Agglutinin; D020106:Acrylamide; D014364:Tryptophan; D014508:Urea; D019791:Guanidine; D001974:Bromosuccinimide