以吉拉德试剂T作为NMR探针研究人源细胞色素c氧化修饰的影响因素
收稿日期: 2025-04-27
网络出版日期: 2025-05-12
基金资助
湖北省自然科学基金创新群体项目(2023AFA041)
Investigating the Factors Influencing Oxidative Modification of Human Cytochrome c Using Girard's Reagent T as an NMR Probe
Received date: 2025-04-27
Online published: 2025-05-12
细胞色素c（Cytochrome c，Cyt c）的氧化修饰可能对蛋白自身的局部构象造成影响，但Cyt c结构的改变如何影响其氧化修饰程度尚不清楚. 本文以吉拉德试剂T（Girard’s Reagent T，GRT）作为核磁共振（NMR）探针，研究了人源Cyt c在不同条件下的氧化修饰程度. 实验结果表明，通过还原甲基化的方法保护蛋白中的赖氨酸，可以降低蛋白的氧化修饰程度；Cyt c发生部分去折叠后，其氧化修饰程度会升高；Cyt c与心磷脂结合后可以很大程度提高蛋白的氧化修饰程度，但蛋白质聚集等其它因素可能对其氧化修饰有抑制作用.
张广庆 , 占建华 , 肖雄 , 朱勤俊 , 蒋滨 , 刘买利 , 张许 . 以吉拉德试剂T作为NMR探针研究人源细胞色素c氧化修饰的影响因素[J]. 波谱学杂志, 2026 , 43(1) : 37 -45 . DOI: 10.11938/cjmr20253164
Oxidative modification of cytochrome c (Cyt c) may influence the local conformation of protein, yet the mechanism by which structural alterations of Cyt c affect its degree of oxidative modification remains unclear. In this study, Girard’s reagent T (GRT) was employed as a nuclear magnetic resonance (NMR) probe to investigate the oxidative modification levels of human Cyt c under varying environmental conditions. Experimental results demonstrated that protecting lysine residues through reductive methylation effectively reduced protein oxidation. Partial unfolding of Cyt c was found to enhance its oxidative modification, while binding Cyt c with cardiolipin significantly increased the extent of oxidation. Additionally, other factors such as protein aggregation exhibited inhibitory effects on oxidative modification.
Key words： cytochrome c; oxidative modification; Girard’s reagent T; methylation; NMR
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