Cisplatin reacts with the RING finger domain of RNF11 and interferes with the protein functions.

IF 2.9 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Metallomics Pub Date : 2023-04-03 DOI:10.1093/mtomcs/mfad017
Yu Wang, Siming Yuan, Kaiming Cao, Yangzhong Liu
{"title":"Cisplatin reacts with the RING finger domain of RNF11 and interferes with the protein functions.","authors":"Yu Wang,&nbsp;Siming Yuan,&nbsp;Kaiming Cao,&nbsp;Yangzhong Liu","doi":"10.1093/mtomcs/mfad017","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":89,"journal":{"name":"Metallomics","volume":"15 4","pages":""},"PeriodicalIF":2.9000,"publicationDate":"2023-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Metallomics","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1093/mtomcs/mfad017","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

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.

顺铂与RNF11的环指结构域发生反应并干扰该蛋白的功能。
蛋白反应在顺铂的作用机制中起重要作用。在这项工作中,我们发现顺铂对参与肿瘤发生和转移的关键蛋白RNF11的环指结构域具有高度反应。结果表明,顺铂在锌配位位点与RNF11结合并导致锌从蛋白质中排出。利用锌染料和硫醇剂进行紫外-可见光谱分析,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,硫醇基团的含量降低。电喷雾电离-质谱测量表明,每个RNF11可以结合多达三个铂原子。动力学分析表明,在t1/2 ~ 3小时内RNF11的铂化率合理。CD、核磁共振和凝胶电泳测量表明,顺铂反应导致RNF11的蛋白展开和寡聚化。下拉实验证实,RNF11的铂化干扰了RNF11与UBE2N的蛋白相互作用,这是RNF11功能化的关键步骤。此外,我们发现Cu(I)可以促进RNF11的铂化,这可能导致高铜水平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的锌释放破坏了蛋白质结构并干扰了其功能。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Metallomics
Metallomics 生物-生化与分子生物学
CiteScore
7.00
自引率
5.90%
发文量
87
审稿时长
1 months
期刊介绍: Global approaches to metals in the biosciences
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信