title: Evidence of stable monomeric species in the unfolding of Cu,Zn superoxide dismutase from Photobacterium leiognathi.
abstract: The equilibrium unfolding process of Photobacterium leiognathi Cu,Zn superoxide dismutase has been quantitatively monitored through circular dichroism (CD) and fluorescence spectroscopy, upon increasing the guanidinium hydrochloride concentration. The study has been undertaken for both the holo- and the copper-free derivative to work out the role of copper in protein stability. In both cases the unfolding was reversible. The denaturation curve derived from CD and fluorescence spectroscopy was not coincident, suggesting that the denaturation process occurs through a three-state model with formation of an intermediate monomeric species. The occurrence of an intermediate species has been unambiguously demonstrated following CD and steady-state fluorescence spectra of the enzyme at various concentrations in presence of a fixed amounts of guanidinium hydrochloride.
authors.affiliation: INFM and Department of Experimental Medicine and Biochemical Sciences, University of Rome "Tor Vergata,", Rome, 00133, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Malvezzi-Campeggi
authors.forename: M E
authors.initials: ME
authors.lastname: Stroppolo
authors.forename: G
authors.initials: G
authors.lastname: Mei
authors.forename: N
authors.initials: N
authors.lastname: Rosato
authors.forename: A
authors.initials: A
authors.lastname: Desideri
mesh_terms: D002942:Circular Dichroism; D019281:Dimerization; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D019791:Guanidine; D020035:Holoenzymes / Q000737:chemistry; D008956:Models, Chemical; D010776:Photobacterium / Q000201:enzymology* / Q000235:genetics; D011489:Protein Denaturation; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013050:Spectrometry, Fluorescence; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000235:genetics; D013816:Thermodynamics
chemical_list: D020035:Holoenzymes; D011994:Recombinant Proteins; D013482:Superoxide Dismutase; D019791:Guanidine