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相关概念视频

Riboswitches01:56

Riboswitches

Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
Transcriptional Regulation: Riboswitches01:23

Transcriptional Regulation: Riboswitches

Riboswitches are RNA elements that regulate gene expression by altering their secondary structures in response to specific effector molecules. These elements, located in the leader regions of certain mRNAs, act as transcriptional regulators by toggling between alternative conformations to control downstream gene expression. Riboswitch-mediated regulation is a precise mechanism for modulating biosynthetic pathways, as exemplified by the riboflavin biosynthesis pathway in Bacillus...
RNA Polymerase II Accessory Proteins02:36

RNA Polymerase II Accessory Proteins

Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
RNA Polymerase II Accessory Proteins02:36

RNA Polymerase II Accessory Proteins

Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
tRNA Activation02:26

tRNA Activation

Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
tRNA Activation02:26

tRNA Activation

Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...

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相关实验视频

Updated: Jun 10, 2026

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases
10:24

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases

Published on: June 30, 2019

激活Ib类核糖核酸减少酶的结构基础

Amie K Boal1, Joseph A Cotruvo, JoAnne Stubbe

  • 1Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, IL 60208, USA.

Science (New York, N.Y.)
|August 7, 2010
PubMed
概括

大肠杆菌中的Ib类核糖核酸减少酶使用或铁辅因子. 结构研究揭示了不同的激活通路和一个关键合因子组装的通道,涉及NrdI蛋白.

科学领域:

  • 生物化学 生物化学
  • 结构生物学 结构生物学
  • 酶学 是一种酶学.

背景情况:

  • 来自大肠杆菌的Ib类核糖核酸减少酶 (RNR) 使用Mn(III) 2-铁基 (Y•) 或Fe(III) 2-Y•辅因子进行核酸减少.
  • 铁辅因子组合与O2是自发的,但辅因子激活需要减少的黄蛋白NrdI.

研究的目的:

  • 阐明了Ib类RNR中辅因子激活的基础结构机制.
  • 调查NrdI在辅因子组装中的作用.

主要方法:

  • 大肠杆菌的X射线晶体学 Mn(II) 2-NrdF和Fe(II) 2-NrdF.
  • 结构分析Mn(II) 2-NrdF与减少和氧化NrdI.F复合体中的结构分析.
  • 反应中间体的结晶学检测.

主要成果:

  • 对于Mn{II}2-NrdF和Fe{II}2-NrdF,观察到不同的协调环境,表明不同的氧化剂结合点.
  • 确定了一条连续通道,将NrdI素辅因子连接到Mn(II) 2-NrdF中的Mn(II) 2-NrdF的活性位点.
  • 在这个通道内检测到一种假定的过氧化物中间体,支持拟议的催化机制.

结论:

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High-throughput Purification of Affinity-tagged Recombinant Proteins
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Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis
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Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis

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Last Updated: Jun 10, 2026

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases
10:24

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases

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High-throughput Purification of Affinity-tagged Recombinant Proteins

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Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis
09:04

Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis

Published on: July 26, 2018

  • 这些发现揭示了和铁辅因子在Ib类RNR中激活的独特结构途径.
  • 通过NrdI介导的辅因子组合包括一个直接通道,促进氧化剂的转移和中间体的形成.