title: Conformational change of the dimeric DsbC molecule induced by GdnHCl. A study by intrinsic fluorescence.
abstract: Unfolding-refolding of Escherichia coli DsbC, a homodimeric molecule, induced by GdnHCl was studied by intrinsic fluorescence. Interpretation of experimental fluorescence data was done together with the analysis of protein 3D structure. It is shown that although Cys 141 is the next neighbor of the single tryptophan residue (Trp 140), the sulfur atoms of the disulfide bond Cys 141-Cys 163 are far apart from the indole ring and cannot quench its fluorescence, while the potential quenchers are Met 136 and His 170. It was revealed that though each subunit of DsbC contains eight tyrosine residues, only three tyrosine residues (Tyr 171, Tyr 38, and Tyr 52) contribute to the bulk fluorescence of the molecule. The character of intrinsic fluorescence intensity changes induced by GdnHCl (equilibrium and kinetic data) and its parametric representation, the existence of an isosbestic point of fluorescence spectra at different GdnHCl concentrations, allowed suggesting a one-step character of DsbC denaturation and its reversibility.
authors.affiliation: Institute of Cytology, Russian Academy of Science, Tihkoretsky av. 4, St. Petersburg 194064, Russia.
authors.forename: Olga V
authors.initials: OV
authors.lastname: Stepanenko
authors.forename: Irina M
authors.initials: IM
authors.lastname: Kuznetsova
authors.forename: Konstantin K
authors.initials: KK
authors.lastname: Turoverov
authors.forename: Chunjuan
authors.initials: C
authors.lastname: Huang
authors.forename: Chih-chen
authors.initials: CC
authors.lastname: Wang
mesh_terms: D019281:Dimerization; D029968:Escherichia coli Proteins / Q000737:chemistry*; D019791:Guanidine / Q000737:chemistry*; D007700:Kinetics; D008956:Models, Chemical; D008958:Models, Molecular; D011487:Protein Conformation; D011489:Protein Denaturation; D019704:Protein Disulfide-Isomerases / Q000737:chemistry*; D017510:Protein Folding; D013050:Spectrometry, Fluorescence / Q000379:methods; D014364:Tryptophan / Q000737:chemistry; D014443:Tyrosine / Q000737:chemistry
chemical_list: D029968:Escherichia coli Proteins; D014443:Tyrosine; D014364:Tryptophan; D019704:Protein Disulfide-Isomerases; D019791:Guanidine