相关实验视频

Updated: Sep 12, 2025

Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations

Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations

Published on: November 7, 2019

描述和设计具有高通量细胞自由表达的翻译后修饰

Derek A Wong^1,2,3, Zachary M Shaver^2,3,4,5, Maria D Cabezas^1,2,3

¹Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, 60208, USA.

Nature communications

|August 5, 2025

概括

一个新的工作流快速测试安装后翻译修改 (PTM) 的蛋白质. 这种方法通过改进设计-构建-测试-学习周期来加速治疗性和糖蛋白的工程.

科学领域:

生物化学生物化学
分子生物学分子生物学
蛋白质工程是指蛋白质工程.

背景情况:

翻译后修改 (PTM) 对治疗性蛋白质和的稳定性和功能至关重要.
现有的研究和工程PTM的方法往往是低吞吐量和实验密集型.

研究的目的:

开发一种可通用的体外工作流程,用于PTM安装蛋白的快速表达和测试.
将此工作流应用于核糖体合成和翻译后修饰的 (RiPPs) 和糖蛋白.
为加速PTM用于治疗应用的工程.

主要方法:

结合无细胞基因表达 (CFE) 与AlphaLISA用于高通量蛋白质分析.
描述RiPP识别元素和整合到生物发现管道中.
调整工作流程以研究和设计用于糖蛋白生产的寡糖糖转移酶 (OST).

主要成果:

证明了用于表征RiPP识别元素绑定活动的工作流程.
成功地将工作流程集成到RiPP产品的生物发现管道中.
鉴定出突变的OST和在模型疫苗载体蛋白上特定的部位,以实现高效的糖化.
启用了高效的糖化蛋白质的生产.

结论:

开发的CFE-AlphaLISA工作流提供了一个快速和可通用的方法来研究和工程PTMs.
这种方法显著加速了蛋白质和治疗工程中的设计-构建-测试-学习循环.
该工作流具有广泛的适用性,特别是对于RiPP和糖蛋白,促进治疗的发展.

更多相关视频

High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes

High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes

Published on: March 3, 2015

High Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension

High Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension

Published on: December 28, 2015

相关概念视频

What is Gene Expression?

What is Gene Expression?

A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then...

Regulation of Expression at Multiple Steps

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...