Banci L, Bertini I, Gallori E, Luchinat C, Paoletti F, Polsinelli M, Viezzoli M S
J Inorg Biochem. 1987 Jun;30(2):77-85. doi: 10.1016/0162-0134(87)80045-4.
The electronic and 1H NMR spectra are reported for the cobalt(II) alkaline phosphatase (EC 3.1.3.1.) system at pH around 6 in the range 0-2 mol of cobalt per mol protein. It is shown that under the present experimental conditions cobalt(II) selectively populates the A sites. Three isotropically shifted NH signals have been detected in the A site that indicate the presence of three histidines in the coordination sphere of cobalt(II). The electronic spectra and the nuclear relaxation properties are consistent with pentacoordination of cobalt(II) in the A site. The finding of reproducible preparation routes for the derivatives, and of appropriate experimental conditions for the observation of their 1H NMR spectra, open new possibilities for the spectroscopic investigation of alkaline phosphatase.
报道了在pH约为6、每摩尔蛋白质含0 - 2摩尔钴的范围内，钴（II）碱性磷酸酶（EC 3.1.3.1.）体系的电子光谱和1H NMR光谱。结果表明，在当前实验条件下，钴（II）选择性地占据A位点。在A位点检测到三个各向同性位移的NH信号，这表明在钴（II）的配位球中有三个组氨酸。电子光谱和核弛豫特性与钴（II）在A位点的五配位一致。衍生物可重复制备路线的发现以及观察其1H NMR光谱的合适实验条件的确定，为碱性磷酸酶的光谱研究开辟了新的可能性。
