Resolution of multiple heme centers of hydroxylamine oxidoreductase from Nitrosomonas. 2. Mössbauer spectroscopy.
Resolution of multiple heme centers of hydroxylamine oxidoreductase from Nitrosomonas. 2. Mössbauer spectroscopy.
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亚硝化单胞菌羟胺氧化还原酶的多个血红素中心的分离。
DOI：
10.1021/bi00260a011
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发表时间：
1982
期刊：
影响因子：
2.9
通讯作者：
Hooper,AB
中科院分区：
文献类型：
--
作者：
Lipscomb,JD;Andersson,KK;Münck,E;Kent,TA;Hooper,AB
材料和方法如前所述从欧洲亚硝化单胞菌的细胞提取物制备HAO（Hooper等人，1978年）。将来自几种制备物的酶合并，并用于如前一篇论文（Lipscomb & Hooper，1982）中所述的EPR研究以及本文所述的穆斯堡尔研究。将酶以冻干粉末的形式储存在-20 ℃下，并重悬于最小体积的50 mM磷酸钾缓冲液（pH7.5）中，用于光谱研究。从光学吸收光谱估计的穆斯堡尔样品中HAO的浓度约为0.3 mM（7.2 mM血红素）。制备一个穆斯堡尔样品所需的纯化酶量需要大约6个月。因此，所有结果都是指对同一样品进行的测量。在氩气气氛下通过加入1重量%的等分试样来进行样品的还原。
Materials and Methods HAO was prepared from cell extracts of Nitrosomonas europaea as previously described (Hooper et al., 1978). The enzyme from several preparations was pooled and used for the EPR studies as described in the preceding paper (Lipscomb & Hooper, 1982) as well as the Mossbauer study described here. The enzyme was stored at-20 C as a lyophilized powder and resuspended in a minimum volume of 50 mM potassium phosphate buffer, pH 7.5, for the spectroscopic studies. The concentration of HAO in the Mossbauersample estimated from the optical absorption spectrum was approximately 0.3 mM (7.2 mM heme). Approximately 6 months was required to prepare the amount of purified enzyme re-quired for a single Mossbauer sample. Consequently, all of the results refer to measurements made on the same sample. Reduction of the sample was performed under an argon atmosphere by adding an aliquot of 1