• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

tRNA 硫醇化酶 TtuA 中 [3Fe-4S] 和 [4Fe-4S] 铁硫簇的快速和自发转化。

Quick and Spontaneous Transformation between [3Fe-4S] and [4Fe-4S] Iron-Sulfur Clusters in the tRNA-Thiolation Enzyme TtuA.

机构信息

Graduate School of Life Science, Hokkaido University, Kita 8, Nishi 5, Kita-ku, Sapporo 060-0810, Japan.

Faculty of Advanced Life Science, Hokkaido University, Kita 8, Nishi 5, Kita-ku, Sapporo 060-0810, Japan.

出版信息

Int J Mol Sci. 2023 Jan 3;24(1):833. doi: 10.3390/ijms24010833.

DOI:10.3390/ijms24010833
PMID:36614280
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9821441/
Abstract

Iron-sulfur (Fe-S) clusters are essential cofactors for enzyme activity. These Fe-S clusters are present in structurally diverse forms, including [4Fe-4S] and [3Fe-4S]. Type-identification of the Fe-S cluster is indispensable in understanding the catalytic mechanism of enzymes. However, identifying [4Fe-4S] and [3Fe-4S] clusters in particular is challenging because of their rapid transformation in response to oxidation-reduction events. In this study, we focused on the relationship between the Fe-S cluster type and the catalytic activity of a tRNA-thiolation enzyme (TtuA). We reconstituted [4Fe-4S]-TtuA, prepared [3Fe-4S]-TtuA by oxidizing [4Fe-4S]-TtuA under strictly anaerobic conditions, and then observed changes in the Fe-S clusters in the samples and the enzymatic activity in the time-course experiments. Electron paramagnetic resonance analysis revealed that [3Fe-4S]-TtuA spontaneously transforms into [4Fe-4S]-TtuA in minutes to one hour without an additional free Fe source in the solution. Although the TtuA immediately after oxidation of [4Fe-4S]-TtuA was inactive [3Fe-4S]-TtuA, its activity recovered to a significant level compared to [4Fe-4S]-TtuA after one hour, corresponding to an increase of [4Fe-4S]-TtuA in the solution. Our findings reveal that [3Fe-4S]-TtuA is highly inactive and unstable. Moreover, time-course analysis of structural changes and activity under strictly anaerobic conditions further unraveled the Fe-S cluster type used by the tRNA-thiolation enzyme.

摘要

铁硫 (Fe-S) 簇是酶活性所必需的辅因子。这些 Fe-S 簇以结构多样的形式存在，包括 [4Fe-4S] 和 [3Fe-4S]。确定 Fe-S 簇的类型对于理解酶的催化机制是不可或缺的。然而，由于它们在氧化还原事件的作用下迅速转化，因此特别难以识别 [4Fe-4S] 和 [3Fe-4S] 簇。在这项研究中，我们专注于 Fe-S 簇类型与 tRNA 硫醇化酶 (TtuA) 的催化活性之间的关系。我们重新构成了 [4Fe-4S]-TtuA，通过在严格的厌氧条件下氧化 [4Fe-4S]-TtuA 制备了 [3Fe-4S]-TtuA，然后观察了样品中 Fe-S 簇的变化和时间进程实验中的酶活性。电子顺磁共振分析表明，[3Fe-4S]-TtuA 在没有溶液中额外游离 Fe 源的情况下，在数分钟到一小时内自发转化为 [4Fe-4S]-TtuA。尽管 [4Fe-4S]-TtuA 氧化后立即形成无活性的 [3Fe-4S]-TtuA，但在一小时后其活性恢复到与 [4Fe-4S]-TtuA 相当的显著水平，这对应于溶液中 [4Fe-4S]-TtuA 的增加。我们的发现表明 [3Fe-4S]-TtuA 高度失活且不稳定。此外，在严格的厌氧条件下进行的结构变化和活性的时程分析进一步揭示了 tRNA 硫醇化酶使用的 Fe-S 簇类型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9bb/9821441/f5a04305055b/ijms-24-00833-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9bb/9821441/9791bc400769/ijms-24-00833-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9bb/9821441/19fc0fc8e09f/ijms-24-00833-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9bb/9821441/39c5bdf082fa/ijms-24-00833-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9bb/9821441/f5a04305055b/ijms-24-00833-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9bb/9821441/9791bc400769/ijms-24-00833-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9bb/9821441/19fc0fc8e09f/ijms-24-00833-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9bb/9821441/39c5bdf082fa/ijms-24-00833-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9bb/9821441/f5a04305055b/ijms-24-00833-g004.jpg

相似文献

1
Quick and Spontaneous Transformation between [3Fe-4S] and [4Fe-4S] Iron-Sulfur Clusters in the tRNA-Thiolation Enzyme TtuA.tRNA 硫醇化酶 TtuA 中 [3Fe-4S] 和 [4Fe-4S] 铁硫簇的快速和自发转化。
Int J Mol Sci. 2023 Jan 3;24(1):833. doi: 10.3390/ijms24010833.
2
Biochemical and structural characterization of oxygen-sensitive 2-thiouridine synthesis catalyzed by an iron-sulfur protein TtuA.铁硫蛋白TtuA催化的氧敏感型2-硫代尿苷合成的生化及结构特征
Proc Natl Acad Sci U S A. 2017 May 9;114(19):4954-4959. doi: 10.1073/pnas.1615585114. Epub 2017 Apr 24.
3
Electron paramagnetic resonance evidence for a novel interconversion of [3Fe-4S](+) and [4Fe-4S](+) clusters with endogenous iron and sulfide in anaerobic ribonucleotide reductase activase in vitro.体外厌氧核糖核苷酸还原酶激活酶中[3Fe-4S](+)和[4Fe-4S](+)簇与内源性铁和硫化物发生新型相互转化的电子顺磁共振证据。
J Biol Chem. 2000 Apr 28;275(17):12367-73. doi: 10.1074/jbc.275.17.12367.
4
The [4Fe-4S] cluster of sulfurtransferase TtuA desulfurizes TtuB during tRNA modification in Thermus thermophilus.硫转移酶 TtuA 的 [4Fe-4S] 簇在嗜热栖热菌中 tRNA 修饰过程中使 TtuB 脱硫。
Commun Biol. 2020 Apr 7;3(1):168. doi: 10.1038/s42003-020-0895-3.
5
Nonredox thiolation in tRNA occurring via sulfur activation by a [4Fe-4S] cluster.通过 [4Fe-4S] 簇的硫原子激活作用发生在 tRNA 中的非氧化型硫醇化反应。
Proc Natl Acad Sci U S A. 2017 Jul 11;114(28):7355-7360. doi: 10.1073/pnas.1700902114. Epub 2017 Jun 27.
6
Effect of cysteine to serine mutations on the properties of the [4Fe-4S] center in Escherichia coli fumarate reductase.半胱氨酸到丝氨酸突变对大肠杆菌延胡索酸还原酶中[4Fe-4S]中心性质的影响。
Biochemistry. 1995 Sep 26;34(38):12284-93. doi: 10.1021/bi00038a024.
7
Redox intermediates of Desulfovibrio gigas [NiFe] hydrogenase generated under hydrogen. Mössbauer and EPR characterization of the metal centers.巨大脱硫弧菌[NiFe]氢化酶在氢气存在下生成的氧化还原中间体。金属中心的穆斯堡尔谱和电子顺磁共振表征。
J Biol Chem. 1989 Oct 5;264(28):16435-50.
8
[3Fe-4S] to [4Fe-4S] cluster conversion in Desulfovibrio fructosovorans [NiFe] hydrogenase by site-directed mutagenesis.通过定点诱变实现果糖脱硫弧菌[NiFe]氢化酶中[3Fe-4S]到[4Fe-4S]簇的转化
Proc Natl Acad Sci U S A. 1998 Sep 29;95(20):11625-30. doi: 10.1073/pnas.95.20.11625.
9
Iron-sulfur stoichiometry and structure of iron-sulfur clusters in three-iron proteins: evidence for [3Fe-4S] clusters.三铁蛋白中铁硫化学计量比及铁硫簇结构：[3Fe-4S]簇的证据
Proc Natl Acad Sci U S A. 1983 Jan;80(2):393-6. doi: 10.1073/pnas.80.2.393.
10
Characterization of the flavins and the iron-sulfur centers of glutamate synthase from Azospirillum brasilense by absorption, circular dichroism, and electron paramagnetic resonance spectroscopies.利用吸收光谱、圆二色光谱和电子顺磁共振光谱对巴西固氮螺菌谷氨酸合酶的黄素和铁硫中心进行表征。
Biochemistry. 1992 May 19;31(19):4613-23. doi: 10.1021/bi00134a011.

引用本文的文献

1
Ferredoxins: Functions, Evolution, Potential Applications, and Challenges of Subtype Classification.铁氧化还原蛋白：功能、进化、潜在应用及亚型分类的挑战
Curr Issues Mol Biol. 2024 Sep 1;46(9):9659-9673. doi: 10.3390/cimb46090574.

本文引用的文献

1
Identification of Iron-Sulfur (Fe-S) Cluster and Zinc (Zn) Binding Sites Within Proteomes Predicted by DeepMind's AlphaFold2 Program Dramatically Expands the Metalloproteome.通过DeepMind的AlphaFold2程序预测蛋白质组中铁硫（Fe-S）簇和锌（Zn）结合位点，极大地扩展了金属蛋白质组。
J Mol Biol. 2022 Jan 30;434(2):167377. doi: 10.1016/j.jmb.2021.167377. Epub 2021 Nov 24.
2
Biosynthesis and Degradation of Sulfur Modifications in tRNAs.tRNA 中硫修饰的生物合成与降解。
Int J Mol Sci. 2021 Nov 3;22(21):11937. doi: 10.3390/ijms222111937.
3
Highly accurate protein structure prediction with AlphaFold.
利用 AlphaFold 进行高精度蛋白质结构预测。
Nature. 2021 Aug;596(7873):583-589. doi: 10.1038/s41586-021-03819-2. Epub 2021 Jul 15.
4
Fe-S cofactors in the SARS-CoV-2 RNA-dependent RNA polymerase are potential antiviral targets.SARS-CoV-2 依赖 RNA 的 RNA 聚合酶中的 Fe-S 辅助因子是潜在的抗病毒靶点。
Science. 2021 Jul 9;373(6551):236-241. doi: 10.1126/science.abi5224. Epub 2021 Jun 3.
5
Iron-sulfur biology invades tRNA modification: the case of U34 sulfuration.铁硫生物学入侵 tRNA 修饰：以 U34 硫化为案例。
Nucleic Acids Res. 2021 Apr 19;49(7):3997-4007. doi: 10.1093/nar/gkab138.
6
Structure-based mechanistic insights into catalysis by tRNA thiolation enzymes.基于结构的tRNA硫醇化酶催化作用的机制性见解。
Curr Opin Struct Biol. 2020 Dec;65:69-78. doi: 10.1016/j.sbi.2020.06.002. Epub 2020 Jul 8.
7
Role of RNA modifications in cancer.RNA 修饰在癌症中的作用。
Nat Rev Cancer. 2020 Jun;20(6):303-322. doi: 10.1038/s41568-020-0253-2. Epub 2020 Apr 16.
8
The [4Fe-4S] cluster of sulfurtransferase TtuA desulfurizes TtuB during tRNA modification in Thermus thermophilus.硫转移酶 TtuA 的 [4Fe-4S] 簇在嗜热栖热菌中 tRNA 修饰过程中使 TtuB 脱硫。
Commun Biol. 2020 Apr 7;3(1):168. doi: 10.1038/s42003-020-0895-3.
9
An ancient type of MnmA protein is an iron-sulfur cluster-dependent sulfurtransferase for tRNA anticodons.一种古老类型的 MnmA 蛋白是一种依赖铁硫簇的 tRNA 反密码子硫转移酶。
RNA. 2020 Mar;26(3):240-250. doi: 10.1261/rna.072066.119. Epub 2019 Dec 4.
10
Recent Advances in Our Understanding of the Biosynthesis of Sulfur Modifications in tRNAs.我们对tRNA中硫修饰生物合成理解的最新进展
Front Microbiol. 2018 Nov 1;9:2679. doi: 10.3389/fmicb.2018.02679. eCollection 2018.