Towards a protocol for solution structure determination of copper(II) proteins: the case of Cu(II)Zn(II) superoxide dismutase.
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.
Bertini
Ivano
I
CERM and Department of Chemistry, University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, Italy. ivanobertini@cerm.unifi.it
Felli
Isabella C
IC
Luchinat
Claudio
C
Parigi
Giacomo
G
Pierattelli
Roberta
R
D003300:Copper / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D011506:Proteins; D011522:Protons; D013482:Superoxide Dismutase / Q000737:chemistry; D015032:Zinc / Q000737:chemistry
D011506:Proteins; D011522:Protons; D003300:Copper; D013482:Superoxide Dismutase; D015032:Zinc