'Light up' protein-protein interaction through bioorthogonal incorporation of a turn-on fluorescent probe into beta-lactamase

	'Light up' protein-protein interaction through bioorthogonal incorporation of a turn-on fluorescent probe into beta-lactamase
	Hu, Rui 1,2,3; Yap, Hong-Kin 1,2; Fung, Yik-Hong 1,2; Wang, Yong 1,2; Cheong, Wing-Lam 1,2; So, Lok-Yan 1,2; Tsang, Chui-Shan 1,2; Lee, Lawrence Yoon Suk 1,2; Lo, Warrick Ken Cheung 1,2; Yuan, Jian 1,2
刊名	MOLECULAR BIOSYSTEMS
	2016
卷号	12 期号:12 页码:3544-3549
英文摘要	Fluorescent labeling of biomacromolecules to 'light up' biological events through non-invasive methods is of great importance, but is still challenging in terms of fluorophore properties and the labeling methods used. Herein, we designed and synthesized a biocompatible and conformation sensitive tetraphenylethene derivative EPB with aggregation induced emission (AIE) properties. By introducing EPB into TEM-1 beta-lactamase (TEM-1 Bla) through a two-step approach, a conformation-dependent fluorescent sensor EPB104-Bla was genetically engineered, which was applied to monitor the proteinprotein interaction (PPI) with beta-lactamase inhibitor protein (BLIP). The fluorescence signal of EPB104-Bla increases by an approximately 5-fold upon binding to BLIP, indicating that EPB-104 Bla is capable of lighting up the PPI. The dissociation constant (K-d) between EPB104-Bla and BLIP was estimated to be 0.6 mu M, which is consistent with that derived from the kinetic inhibition assay. This study demonstrates that genetic modification of proteins with AIE probes might open up new opportunities to develop biosensors in PPI analysis.
收录类别	SCI
语种	英语
内容类型	期刊论文
源URL	[http://ir.iccas.ac.cn/handle/121111/35694]
专题	化学研究所_光化学实验室
作者单位	1.Hong Kong Polytech Univ, Dept Appl Biol & Chem Technol, Kowloon, Hong Kong, Peoples R China 2.Hong Kong Polytech Univ, State Key Lab Chirosci, Kowloon, Hong Kong, Peoples R China 3.Chinese Acad Sci, BNLMS, Inst Chem, Key Lab Photochem, Beijing 100190, Peoples R China
推荐引用方式 GB/T 7714	Hu, Rui,Yap, Hong-Kin,Fung, Yik-Hong,et al. 'Light up' protein-protein interaction through bioorthogonal incorporation of a turn-on fluorescent probe into beta-lactamase[J]. MOLECULAR BIOSYSTEMS,2016,12(12):3544-3549.
APA	Hu, Rui.,Yap, Hong-Kin.,Fung, Yik-Hong.,Wang, Yong.,Cheong, Wing-Lam.,...&Wong, Kwok-Yin.(2016).'Light up' protein-protein interaction through bioorthogonal incorporation of a turn-on fluorescent probe into beta-lactamase.MOLECULAR BIOSYSTEMS,12(12),3544-3549.
MLA	Hu, Rui,et al."'Light up' protein-protein interaction through bioorthogonal incorporation of a turn-on fluorescent probe into beta-lactamase".MOLECULAR BIOSYSTEMS 12.12(2016):3544-3549.