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本实验首次以稳定修饰试剂[PtdienNO_3]Cl标记细胞色素c,反应产物经CM-52阳离子交换色谱分离纯化得细胞色素c单修饰、双修饰及三修饰衍生物。用微分脉冲伏安测定各修饰产物的还原电位,并以~1H NMR波谱证实[PtdienNO_3]Cl在细胞色素c上的修饰位点为His-33和Trp-59,其中Trp-59是其它过渡金属配合物所未能标记的、与血红素以氢键相连的保守氨基酸。细胞色素c Trp-59标记衍生物的获得,为研究芳香族氨基酸在细胞色素c电子转移过程中的作用提供了有价值的实验材料。
In this experiment, cytochrome c was labeled with stable modified reagent [PtdienNO_3] Cl for the first time. The reaction products were separated and purified by CM-52 cation exchange chromatography to obtain single, double and triple modified derivatives of cytochrome c. The reduction potential of each modified product was determined by differential pulse voltammetry. The sites of [PtdienNO_3] Cl modification on cytochrome c were confirmed to be His-33 and Trp-59 by ~ 1H NMR spectroscopy. Among them, Trp-59 was the most abundant transition metal Unlabeled complexes, with hydrogen bonds with hydrogen bonds of the conservative amino acids. Cytochrome c Trp-59 labeled derivatives obtained for the study of aromatic amino acids in the cytochrome c electron transfer process provides valuable experimental materials.