根据反应性分析的mRNA显示,准确地广播乳生物合成途径的基质适应性
Alexander A Vinogradov1, Emiko Nagai2, Jun Shi Chang1
1Department of Chemistry, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Journal of the American Chemical Society
|November 19, 2020
概括
我们开发了一种高通量方法,用于对核糖体合成和转化后修饰 (RiPP) 生物合成的酶反应. 这种技术准确地绘制了酶基质偏好,有助于发现新.
科学领域:
- 生物化学
- 分子生物学
- 合成生物学
背景情况:
- 核糖体合成和转化后改性 (RiPPs) 是一种具有复杂生物合成途径的多样性自然产品.
- 了解RiPP生物合成酶的基质特异性对于阐明这些途径和设计新型来说至关重要.
研究的目的:
- 开发和验证一种高吞吐量方法来分析RiPP生物合成酶的反应性.
- 研究乳糖醇生物合成途径的基质适应性.
- 为发现具有设计生物活动的新型树脂奠定基础.
主要方法:
- 使用mRNA显示技术创建乳前体的和突变性库.
- 应用高通量反应性分析以分析酶基质偏好.
- 通过体外测试获得量化反应和时间分辨率.
主要成果:
- 开发的试验证明了 in vitro 数据的准确性和可重复性.
- 成功地回顾了已知的乳生物合成方面.
- 通过识别乳生物合成酶的基质杂交,扩大了知识.
结论:
- 高通量反应性分析方法是研究RiPP酶基质适应性景观的强大工具.
- 这种方法有助于快速分析酶偏好和发现新型RiPP.
- 这些发现支持组合性库的构建,用于新的设计.
相关概念视频
From DNA to Protein
The flow of genetic information in cells from DNA to mRNA to protein is described by the central dogma, which states that genes specify the sequence of mRNAs, which in turn specify the sequence of amino acids making up all proteins. The decoding of one molecule to another is performed by specific proteins and RNAs. Because the information stored in DNA is so central to cellular function, it makes intuitive sense that the cell would make mRNA copies of this information for protein synthesis...
Leaky Scanning
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA. Marilyn Kozak discovered that the sequence RCCAUGG (where R stands for...
Ribosome Profiling
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
mRNA Stability and Gene Expression
The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
Cis-acting Elements involved in mRNA stability
Regulation of Expression at Multiple Steps
The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the addition of a...
Nucleic Acid Structure
The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group on the ribose's second carbon and a hydrogen on the deoxyribose's second carbon. The phosphate residue attaches to the hydroxyl group of the 5′ carbon of one sugar and the hydroxyl group of the 3′ carbon of the sugar of the next nucleotide, which forms a 5′ to 3′ phosphodiester linkage.
DNA Structure
DNA has a double-helix structure. The...
DNA Structure
DNA has a double-helix structure. The...


