The solution structure of the demetalated copper, zinc superoxide dismutase is obtained for the monomeric Glu133Gln/Phe50Glu/Gly51Glu mutant through NMR spectroscopy. The demetalated protein still has a well-defined tertiary structure; however, two β-strands containing two copper ligands (His46 and His48, β4) and one zinc ligand (Asp83, β5) are shortened, and the sheet formed by these strands and strands β7 and β8 moves away from the other strands of the β-barrel to form an open clam with respect to a closed conformation in the holoprotein. Furthermore, loop IV which contains three zinc ligands (His63, His71, and His80) and loop VII which contributes to the definition of the active cavity channel are severely disordered, and experience extensive mobility as it results from thorough 15N relaxation measurements. These structural and mobility data, if compared with those of the copper-depleted protein and holoprotein, point out the role of each metal ion in the protein folding, leading to the final tertiary structure of the holoprotein, and provide hints for the mechanisms of metal delivery by metal chaperones.
Solution structure of apo Cu,Zn superoxide dismutase: the role of metal ions in protein folding / L. BANCI; I. BERTINI; F. CRAMARO; R. DEL CONTE; M.S. VIEZZOLI. - In: BIOCHEMISTRY. - ISSN 0006-2960. - STAMPA. - 42:(2003), pp. 9543-9553. [10.1021/bi034324m]
Solution structure of apo Cu,Zn superoxide dismutase: the role of metal ions in protein folding
BANCI, LUCIA;BERTINI, IVANO;DEL CONTE, REBECCA;VIEZZOLI, MARIA SILVIA
2003
Abstract
The solution structure of the demetalated copper, zinc superoxide dismutase is obtained for the monomeric Glu133Gln/Phe50Glu/Gly51Glu mutant through NMR spectroscopy. The demetalated protein still has a well-defined tertiary structure; however, two β-strands containing two copper ligands (His46 and His48, β4) and one zinc ligand (Asp83, β5) are shortened, and the sheet formed by these strands and strands β7 and β8 moves away from the other strands of the β-barrel to form an open clam with respect to a closed conformation in the holoprotein. Furthermore, loop IV which contains three zinc ligands (His63, His71, and His80) and loop VII which contributes to the definition of the active cavity channel are severely disordered, and experience extensive mobility as it results from thorough 15N relaxation measurements. These structural and mobility data, if compared with those of the copper-depleted protein and holoprotein, point out the role of each metal ion in the protein folding, leading to the final tertiary structure of the holoprotein, and provide hints for the mechanisms of metal delivery by metal chaperones.
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