We have developed an optimized protocol to solve the solution structure of copper(II) proteins. After assignment, proton–proton NOEs are used for the shell where 1H spectra are conveniently observed. In a shell closer to the metal ion, 13C NMR spectra with band-selective homonuclear decoupling provide the assignment of all nuclei except for those of the metal ligands. A convenient method for the measurement of 13C longitudinal-relaxation rates (R1) of carbonyls and carboxylate moieties is proposed. 1H NOEs and 1H and 13C R1 data are sufficient to produce a good/reasonable solution structure, as demonstrated for a monomeric species of superoxide dismutase, a 153-residue protein.
Towards a protocol for solution structure determination of copper(II) proteins: the case of Cu(II)Zn(II) superoxide dismutase / I.Bertini; I.C.Felli; C.Luchinat; G.Parigi; R.Pierattelli. - In: CHEMBIOCHEM. - ISSN 1439-4227. - STAMPA. - 8:(2007), pp. 1422-1429. [10.1002/cbic.200700006]
Towards a protocol for solution structure determination of copper(II) proteins: the case of Cu(II)Zn(II) superoxide dismutase
BERTINI, IVANO;FELLI, ISABELLA CATERINA;LUCHINAT, CLAUDIO;PARIGI, GIACOMO;PIERATTELLI, ROBERTA
2007
Abstract
We have developed an optimized protocol to solve the solution structure of copper(II) proteins. After assignment, proton–proton NOEs are used for the shell where 1H spectra are conveniently observed. In a shell closer to the metal ion, 13C NMR spectra with band-selective homonuclear decoupling provide the assignment of all nuclei except for those of the metal ligands. A convenient method for the measurement of 13C longitudinal-relaxation rates (R1) of carbonyls and carboxylate moieties is proposed. 1H NOEs and 1H and 13C R1 data are sufficient to produce a good/reasonable solution structure, as demonstrated for a monomeric species of superoxide dismutase, a 153-residue protein.
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