1H NMR investigation of cytochrome cd1: complexes with electron-donor proteins.
1H NMR investigation of cytochrome cd1: complexes with electron-donor proteins.
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细胞色素 cd1 的 1 H NMR 研究：与电子供体蛋白的复合物。
DOI：
10.1021/bi00353a022
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发表时间：
1986
期刊：
影响因子：
2.9
通讯作者：
Timkovich,R
中科院分区：
文献类型：
--
作者：
Timkovich,R
材料和方法纯化来自铜绿假单胞菌菌株ATCC 19429的细胞色素cdx、细胞色素c-551和天青蛋白，并如前所述进行动力学测定（Dhesi和Timkovich，1984）。用于NMR研究的蛋白质样品相对于50 mM碳酸氢铵缓冲液（pH7.8）透析，冻干，并作为浓缩的储备溶液再溶解于用20 mM磷酸钾缓冲的99.8%氧化氘中，至约100 mg/ml。所报告的pH值是先前校准的玻璃微电极相对于质子标准物的直接阅读，并且将被指定为pH*。通过紫外-可见分光光度法测定储备蛋白溶液的浓度。报告的细胞色素cd浓度是指亚基浓度。混合等分试样，必要时用缓冲液稀释，得到最终混合物。
Materials and MethodsCytochrome cdx, cytochrome c-551, and azurin from P. aeruginosa strain ATCC 19429 were purified and kinetic assays performed as described previously (Dhesi & Timkovich, 1984). Horse heart cytochrome c, type III, was purchased from Sigma Chemical Co.Protein samples for NMR studies were dialyzed vs. 50 mM ammonium bicarbonate buffer, pH 7.8, lyophilized, and re-dissolved as concentrated stock solutions in 99.8% deuterium oxide buffered with 20 mM potassium phosphate to an ap-propriate pH. Values of pH reported are the direct reading of a glass microelectrode previously calibrated vs. protic standards and will be designated pH*. Concentrations of stock protein solutions were determined by UV-visible spectro-photometry. The reported concentrations of cytochrome cd will refer to the subunit concentration. Aliquots were mixed and diluted with buffer if necessary to give the final mixtures.