Purification and characterization of recombinant Caulobacter crescentus Cu,Zn superoxide dismutase.
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.
De Domenico
Ivana
I
Department of Microbiological, Genetic and Molecular Sciences, University of Messina, Salita Sperone, 31, 98166 Messina, Italy.
Lania
Amalia
A
Bonaccorsi di Patti
Maria Carmela
MC
Battistoni
Andrea
A
Musci
Giovanni
G
Desideri
Alessandro
A
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
D011994:Recombinant Proteins; D013482:Superoxide Dismutase