title: Unfolding of dimeric creatine kinase in urea and guanidine hydrochloride as measured using small angle X-ray scattering with synchrotron radiation.
abstract: The denaturation of dimeric creatine kinase (CK) induced by urea and guanidine hydrochloride (GuHCl) has been studied by small angle X-ray scattering (SAXS), which is a direct way to measure the changes in the overall dimensions of a protein molecule. The radii of gyration (Rg) of CK are 29+/-0.4 angstroms in the native state and 46+/-1.5 angstroms in the unfolded state in either 8 M urea or 3 M GuHCl. The transition curves of urea denaturation derived from the Rg values and the zero angle intensity (I(0)) are similar to that from intrinsic fluorescence, indicating that the changes in the molecular shape, the tertiary structure and the dissociation of the subunits proceed simultaneously. In the case of GuHCl-induced denaturation, the dramatic increases both in Rg and in I(0) in 0.3-0.5 M GuHCl suggest clearly that soluble aggregates form at low GuHCl concentrations. The aggregates dissociate and the molecule unfolds at higher GuHCl concentrations. The results suggest that the mechanisms of CK denaturation in urea and in GuHCl are somewhat different and the intermediate in GuHCl denaturation can much more easily form soluble aggregates.
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Academia Sinica, Beijing, China. zhoujm@sun5.ibp.ac.cn
authors.forename: J M
authors.initials: JM
authors.lastname: Zhou
authors.forename: Y X
authors.initials: YX
authors.lastname: Fan
authors.forename: H
authors.initials: H
authors.lastname: Kihara
authors.forename: K
authors.initials: K
authors.lastname: Kimura
authors.forename: Y
authors.initials: Y
authors.lastname: Amemiya
mesh_terms: D000818:Animals; D003402:Creatine Kinase / Q000737:chemistry*; D019281:Dimerization; D005453:Fluorescence; D019791:Guanidine; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011817:Rabbits; D012542:Scattering, Radiation; D014508:Urea; D014965:X-Rays
chemical_list: D014508:Urea; D003402:Creatine Kinase; D019791:Guanidine