title: Purification and characterization of recombinant Caulobacter crescentus Cu,Zn superoxide dismutase.
abstract: Recombinant Cu,Zn Superoxide Dismutase from Caulobacter crescentus has been expressed in Escherichia coli and characterized. The corresponding recombinant protein has a molecular weight typical of a homodimeric Cu,ZnSODs and an activity comparable to that of other prokaryotic enzymes. The copper active site is characterized by a peculiar axial geometry as evidenced by its electron paramagnetic resonance spectrum, moreover, the copper atom displays a low accessibility toward external chelating agents indicating a lower solvent accessibility when compared to other prokaryotic enzymes. Investigation of the enzyme thermal stability through differential scanning calorimetry indicates the occurrence of two transitions at low and higher temperature that are found to be due to the apo and holo protein, respectively, confirming that the metals have a crucial role in the stabilization of this class of enzymes.
authors.affiliation: Department of Microbiological, Genetic and Molecular Sciences, University of Messina, Salita Sperone, 31, 98166 Messina, Italy.
authors.forename: Ivana
authors.initials: I
authors.lastname: De Domenico
authors.forename: Amalia
authors.initials: A
authors.lastname: Lania
authors.forename: Maria Carmela
authors.initials: MC
authors.lastname: Bonaccorsi di Patti
authors.forename: Andrea
authors.initials: A
authors.lastname: Battistoni
authors.forename: Giovanni
authors.initials: G
authors.lastname: Musci
authors.forename: Alessandro
authors.initials: A
authors.lastname: Desideri
mesh_terms: D000595:Amino Acid Sequence; D016935:Caulobacter crescentus / Q000201:enzymology* / Q000235:genetics; D004578:Electron Spin Resonance Spectroscopy; D007700:Kinetics; D008969:Molecular Sequence Data; D010776:Photobacterium / Q000201:enzymology / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D013816:Thermodynamics
chemical_list: D011994:Recombinant Proteins; D013482:Superoxide Dismutase