title: Evidence for the existence of an unfolding intermediate of thyroglobulin during denaturation by guanidine hydrochloride.
abstract: The unfolding of bovine thyroglobulin (Tg) in guanidine hydrochloride (GuHCl) solution was studied by following the fluorescence and circular dichroism. With increasing GuHCl concentrations, the emission maximum of the intrinsic fluorescence clearly red-shifted in two stages. At concentrations of GuHCl less than 1.2 M or more than 1.6 M, the red shift showed a cooperative manner. At concentrations of GuHCl between 1.2 and 1.6 M, an unfolding intermediate was observed. It was further characterized by the increased binding of the fluorescence probe 1-anilinonaphthalene-8-sulfonic acid (ANS). No significant changes of the secondary structure were indicated by CD spectra at the concentrations of GuHCl between 1.2 and 1.6 M. The conformation of this state has properties similar to those of a molten globule state which may exist in the folding pathway of the protein. Further changes in fluorescence properties occurred at concentrations of denaturant higher than 1.6 M with a significant red shift of the emission maximum from 340 to 347 nm and a marked decrease in ANS binding. This in vitro study gave a clue to understand the biochemical mechanism for the occurrence of aggregation and molecular chaperone binding during Tg maturation in vivo.
authors.affiliation: Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, 100084, China. zhm-dbs@tsinghua.edu.cn
authors.forename: Y-K
authors.initials: YK
authors.lastname: Hong
authors.forename: F-G
authors.initials: FG
authors.lastname: Meng
authors.forename: H
authors.initials: H
authors.lastname: Tang
authors.forename: H-M
authors.initials: HM
authors.lastname: Zhou
mesh_terms: D000817:Anilino Naphthalenesulfonates / Q000737:chemistry; D000818:Animals; D002417:Cattle; D002942:Circular Dichroism; D019791:Guanidine / Q000737:chemistry*; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D013050:Spectrometry, Fluorescence; D013954:Thyroglobulin / Q000737:chemistry*
chemical_list: D000817:Anilino Naphthalenesulfonates; C027132:1-anilino-8-naphthalenesulfonate; D013954:Thyroglobulin; D019791:Guanidine