气候变化领域集成服务门户(CCPortal): An enzyme-based biosensor for monitoring and engineering protein stability in vivo

CCPortal

DOI	10.1073/pnas.2101618118
	An enzyme-based biosensor for monitoring and engineering protein stability in vivo
	Ren C.; Wen X.; Mencius J.; Quan S.
发表日期	2021
ISSN	00278424
卷号	118 期号:13
英文摘要	Protein stability affects the physiological functions of proteins and is also a desirable trait in many protein engineering tasks, yet improving protein stability is challenging because of limitations in methods for directly monitoring protein stability in cells. Here, we report an in vivo stability biosensor wherein a protein of interest (POI) is inserted into a microbial enzyme (CysGA) that catalyzes the formation of endogenous fluorescent compounds, thereby coupling POI stability to simple fluorescence readouts. We demonstrate the utility of the biosensor in directed evolution to obtain stabilized, less aggregation-prone variants of two POIs (including nonamyloidogenic variants of human islet amyloid polypeptide). Beyond engineering applications, we exploited our biosensor in deep mutational scanning for experimental delineation of the stability-related contributions of all residues throughout the catalytic domain of a histone H3K4 methyltransferase, thereby revealing its scientifically informative stability landscape. Thus, our highly accessible method for in vivo monitoring of the stability of diverse proteins will facilitate both basic research and applied protein engineering efforts. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Biosensor; Deep mutational scanning; Protein engineering; Protein stability
语种	英语
scopus关键词	histone methyltransferase; microbial enzyme; Article; catalysis; controlled study; enzyme active site; human; in vivo study; monitoring; mutation; nonhuman; priority journal; protein engineering; protein stability
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/180082
作者单位	State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing, East China University of Science and Technology, Shanghai, 200237, China
推荐引用方式 GB/T 7714	Ren C.,Wen X.,Mencius J.,et al. An enzyme-based biosensor for monitoring and engineering protein stability in vivo[J],2021,118(13).
APA	Ren C.,Wen X.,Mencius J.,&Quan S..(2021).An enzyme-based biosensor for monitoring and engineering protein stability in vivo.Proceedings of the National Academy of Sciences of the United States of America,118(13).
MLA	Ren C.,et al."An enzyme-based biosensor for monitoring and engineering protein stability in vivo".Proceedings of the National Academy of Sciences of the United States of America 118.13(2021).