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. Study holds ProTherm entries: 12864, 12865, 12866, 12867 Extra Details: Holo. dimer <- -> monomer(Intermediate). Transition 1. protein stability; dimeric interaction; protein unfolding; lifetime distributions
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:44 p.m.
|Number of data points||8|
|Proteins||Superoxide dismutase [Cu-Zn] ; SUPEROXIDE DISMUTASE|
|Assays/Quantities/Protocols||Experimental Assay: m ; Experimental Assay: dG_H2O|
|Libraries||Mutations for sequence QDLTVKMTDLQTGKPVGTIELSQNKYGVVFIPELADLTPGMHGFHIHQNGSCASSEKDGKVVLGGAAGGHYDPEHTNKHGFPWTDDNHKGDLPALFVSANGLATNPVLAPRLTLKELKGHAIMIHAGGDNHSDMPKALGGGGARVACGVIQ|