Protein reactions play important roles in the mechanism of action of cisplatin. In this work, we found that cisplatin is highly reactive to the RING finger domain of RNF11, a key protein involved in tumorigenesis and metastasis. The results show that cisplatin binds to RNF11 at the zinc coordination site and leads to zinc ejection from the protein. The formation of S-Pt(II) coordination and Zn(II) ions release have been confirmed by UV-vis spectrometry using zinc dye and thiol agent, showing reducing the contents of thiol groups while forming S-Pt bonds and releasing zinc ions. Electrospray ionization-mass spectrometry measurement indicates that each RNF11 can bind up to three platinum atoms. Kinetical analysis shows a reasonable platination rate of RNF11 with t1/2 ∼ 3 h. CD, nuclear magnetic resonance, and gel electrophoresis measurements indicate that the cisplatin reaction causes protein unfolding and oligomerization of RNF11. Pull-down assay confirms that the platination of RNF11 interferes with the protein interaction of RNF11 with UBE2N, a key step of the functionalization of RNF11. Furthermore, Cu(I) was found to promote the platination of RNF11, which could lead to increased protein reactivity to cisplatin in tumor cells with high copper levels. These results indicate that the platination-induced zinc release of RNF11 disrupts the protein structure and interferes with its functions.

中文翻译：

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顺铂与 RNF11 的环指结构域发生反应并干扰蛋白质功能。

蛋白质反应在顺铂的作用机制中起重要作用。在这项工作中，我们发现顺铂对 RNF11 的环指结构域具有高度反应性，RNF11 是一种参与肿瘤发生和转移的关键蛋白。结果表明，顺铂在锌配位位点与 RNF11 结合，导致锌从蛋白质中排出。使用锌染料和硫醇剂通过紫外-可见光谱法证实了 S-Pt(II) 配位的形成和 Zn(II) 离子的释放，表明在形成 S-Pt 键和释放锌离子的同时降低了硫醇基的含量。电喷雾电离质谱测量表明每个 RNF11 最多可以结合三个铂原子。动力学分析表明 RNF11 的铂化率合理，t1/2 ∼ 3 h。CD、核磁共振、和凝胶电泳测量表明顺铂反应导致 RNF11 的蛋白质去折叠和寡聚化。Pull-down 测定证实 RNF11 的铂化干扰了 RNF11 与 UBE2N 的蛋白质相互作用，这是 RNF11 功能化的关键步骤。此外，发现 Cu(I) 促进 RNF11 的铂化，这可能导致高铜水平的肿瘤细胞中蛋白质对顺铂的反应性增加。这些结果表明，铂化诱导的 RNF11 锌释放破坏了蛋白质结构并干扰了其功能。发现 Cu(I) 促进 RNF11 的铂化，这可能导致高铜水平的肿瘤细胞中蛋白质对顺铂的反应性增加。这些结果表明，铂化诱导的 RNF11 锌释放破坏了蛋白质结构并干扰了其功能。发现 Cu(I) 促进 RNF11 的铂化，这可能导致高铜水平的肿瘤细胞中蛋白质对顺铂的反应性增加。这些结果表明，铂化诱导的 RNF11 锌释放破坏了蛋白质结构并干扰了其功能。