CERM and Department of Chemistry, University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, Italy.
Chembiochem. 2007 Aug 13;8(12):1422-9. doi: 10.1002/cbic.200700006.
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.
我们开发了一种优化的方案来解决铜（II）蛋白的溶液结构问题。在分配之后，质子-质子 NOE 可用于方便观察 1H 谱的壳层。在更接近金属离子的壳层中，带有带选择性同核去耦的 13C NMR 谱可提供除金属配体核之外的所有核的分配。提出了一种测量羰基和羧酸盐部分的 13C 纵向弛豫率（R1）的方便方法。如 1H NOE 和 1H 和 13C R1 数据足以产生良好/合理的溶液结构所示，这对于超氧化物歧化酶的单体物种（一种含有 153 个残基的蛋白质）进行了证明。
