PUP-IT2 as an alternative strategy for PUP-IT proximity labeling

doi:10.3389/fmolb.2022.1007720

	PUP-IT2 as an alternative strategy for PUP-IT proximity labeling
	Yue, Suyu; Xu, Peng; Cao, Zhihe; Zhuang, Min
	2022-09-29
发表期刊	FRONTIERS IN MOLECULAR BIOSCIENCES
ISSN	2296-889X
EISSN	2296-889X
卷号	9
发表状态	已发表
DOI	10.3389/fmolb.2022.1007720
摘要	PUP-IT is a proximity labeling method based on the prokaryotic enzyme PafA. PafA mediates the ligation of Pup, a small peptide, to the proximal proteins. It is different from other proximity labeling methods, such as BioID and APEX, in that both the enzyme and the labeling tag are proteins, which allows for potential in vivo applications. All proximity labeling involves the genetic fusion of the proximity labeling enzyme with the bait protein. However, PafA is a 55 kDa enzyme which sometimes interferes with the bait function. In this study, we tested an alternative proximity labeling strategy, PUP-IT2, in which only a small 7 kDa protein is fused to the bait protein. We examined the activity of PUP-IT2 in vitro and in cells. We also compared it with the original PUP-IT. Finally, we applied PUP-IT2 coupled mass spectrometry to map protein-protein interactions. Overall, we established a new way to use PUP-IT2 for proximity labeling, and this method may have a broad application.
关键词	PUP-IT ligase protein labeling proximity tagging PafA
URL	查看原文
收录类别	SCI ; SCIE
语种	英语
资助项目	Science and Technology Commission of Shanghai Municipality[31922038] ; National Key R&D Program of China[19JC1413700] ; Shanghai Frontiers Science Center for Biomacromolecules and Precision Medicine at ShanghaiTech University["2021YFA0804700","2021YFA1100800"]
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemistry & Molecular Biology
WOS记录号	WOS:000868635100001
出版者	FRONTIERS MEDIA SA
引用统计
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/240469
专题	生命科学与技术学院_PI研究组_庄敏组生命科学与技术学院_硕士生生命科学与技术学院_博士生
通讯作者	Zhuang, Min
作者单位	ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China
第一作者单位	生命科学与技术学院
通讯作者单位	生命科学与技术学院
第一作者的第一单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Yue, Suyu,Xu, Peng,Cao, Zhihe,et al. PUP-IT2 as an alternative strategy for PUP-IT proximity labeling[J]. FRONTIERS IN MOLECULAR BIOSCIENCES,2022,9.
APA	Yue, Suyu,Xu, Peng,Cao, Zhihe,&Zhuang, Min.(2022).PUP-IT2 as an alternative strategy for PUP-IT proximity labeling.FRONTIERS IN MOLECULAR BIOSCIENCES,9.
MLA	Yue, Suyu,et al."PUP-IT2 as an alternative strategy for PUP-IT proximity labeling".FRONTIERS IN MOLECULAR BIOSCIENCES 9(2022).