Pulsed electron paramagnetic resonance studies of the copper(II) site in galactose oxidase.
Pulsed electron paramagnetic resonance studies of the copper(II) site in galactose oxidase.
复制标题
半乳糖氧化酶中铜 (II) 位点的脉冲电子顺磁共振研究。
DOI：
10.1021/bi00548a007
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发表时间：
1980
期刊：
影响因子：
2.9
通讯作者：
W. Mims
中科院分区：
文献类型：
--
作者：
D. Kosman;J. Peisach;W. Mims
材料与方法半乳糖氧化酶（ec1.1）。3.9)按照描述分离（Kosman et al., 1974）。用部分中和的KCN，在pH 8.3的1:1甘油-水混合物中，用光学滴定法制备了铜（II）二乙基三胺氰配合物。咪唑（Sigma化学公司）在用Norite初始处理后由苯重结晶两次。KC15N（99原子%，5N）购自Stohler同位素化学公司。KF是贝尔实验室的区域精制样品。电子自旋回波包络的测量是按照mim & Peisach （1976a）的描述进行的，而LEFE的测量是按照Peisach & Mims（1973）和Mims（1974）的描述进行的。在三脉冲实验中获得的回波包络如前所述进行傅里叶变换（Shimizu et al., 1979; Mims & Peisach, 1979）。
Materials and MethodsGalactose oxidase (EC 1.1. 3.9) was isolated as described (Kosman et al., 1974). Copper (II) diethylenetriamine cyanide complexes were prepared by optical titration of the copper amine complex in a 1: 1 glycerol-H20 mixture at pH 8.3 with partially neutralized KCN. Imidazole (Sigma Chemical Co.) was twice recrystallized from benzene following an initial treatment with Norite. KC15N (99 atom%, 5N) was pur-chased from Stohler Isotope Chemicals. KF was a zone-re-fined sample from Bell Laboratories. Measurements of the electron spin-echo envelope were made as described by Mims & Peisach (1976a), and the LEFE measurements were made as described by Peisach & Mims (1973) and Mims (1974). The echo envelopes obtained in three-pulse experiments were Fourier transformed as described previously (Shimizu et al., 1979; Mims & Peisach, 1979).