相关实验视频

Updated: Jul 24, 2025

Mapping the Binding Site of an Aptamer on ATP Using MicroScale Thermophoresis

Mapping the Binding Site of an Aptamer on ATP Using MicroScale Thermophoresis

Published on: January 7, 2017

原生自上而下的质谱检测揭示了功能性菌素感应的两种不同的结合基因

Sarah Viola Heel¹, Karolina Bartosik¹, Fabian Juen¹

¹Institute of Organic Chemistry and Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.

Journal of the American Chemical Society

|July 7, 2023

概括

通过质谱检测发现Neomycin B在多个位点与RNA核突变器结合. 修改RNA结构改变了neomycin B的结合,突出了非正规基对在氨基糖体识别中的作用.

科学领域:

生物化学
分子生物学
结构生物学

背景情况:

与核糖核酸 (RNA) 的结合对生物过程和药物开发至关重要.
尼奥米辛B是一种氨基糖类抗生素,已知与RNA的相互作用.

研究的目的:

研究neomycin B与neomycin-sensing riboswitch aptamer结构的结合情况.
通过质谱测量确定neomycin B的结合部位和相互作用.

主要方法:

具有电喷离子 (ESI) 和碰撞激活解离 (CAD) 的自主上下质谱 (MS).
对27nt和40nt的aptamer结构进行分析.
与核磁共振 (NMR) 数据进行结构分析和比较.

主要成果:

对27nt构造的MS数据证实了与NMR结构一致的结合点和相互作用.
对于一个40nt的结构,确定了两个同样密集的neomycin B结合基因:一个膨胀环和一个小的沟相互作用.
在下茎的RNA基对修改显著改变了neomycin B的结合亲和力和因子之间的分布.

结论:

质谱检测提供了位点特异的,通过石化测量得到的关于氨基糖类-RNA相互作用的信息.
非正规基因对在RNA识别中发挥着重要作用,例如neomycin B.
了解这些相互作用对于基于RNA的药物开发至关重要.

更多相关视频

An Assay for Quantifying Protein-RNA Binding in Bacteria

An Assay for Quantifying Protein-RNA Binding in Bacteria

Published on: June 12, 2019

A Method for Selecting Structure-switching Aptamers Applied to a Colorimetric Gold Nanoparticle Assay

A Method for Selecting Structure-switching Aptamers Applied to a Colorimetric Gold Nanoparticle Assay

Published on: February 28, 2015

相关概念视频

Riboswitches

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...

Ribosome Profiling

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...