title: 'Light up' protein-protein interaction through bioorthogonal incorporation of a turn-on fluorescent probe into β-lactamase.
abstract: 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 β-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 protein-protein interaction (PPI) with β-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 (Kd) between EPB104-Bla and BLIP was estimated to be 0.6 μM, 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.
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk and Beijing National Laboratory for Molecular Sciences (BNLMS), Key Laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China. gqyang@iccas.ac.cn.
authors.forename: Rui
authors.initials: R
authors.lastname: Hu
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk.
authors.forename: Hong-Kin
authors.initials: HK
authors.lastname: Yap
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk.
authors.forename: Yik-Hong
authors.initials: YH
authors.lastname: Fung
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk.
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk.
authors.forename: Wing-Lam
authors.initials: WL
authors.lastname: Cheong
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk.
authors.forename: Lok-Yan
authors.initials: LY
authors.lastname: So
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk.
authors.forename: Chui-Shan
authors.initials: CS
authors.lastname: Tsang
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk.
authors.forename: Lawrence Yoon Suk
authors.initials: LY
authors.lastname: Lee
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk.
authors.forename: Warrick Ken Cheung
authors.initials: WK
authors.lastname: Lo
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk.
authors.forename: Jian
authors.initials: J
authors.lastname: Yuan
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk.
authors.forename: Ning
authors.initials: N
authors.lastname: Sun
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk.
authors.forename: Yun-Chung
authors.initials: YC
authors.lastname: Leung
authors.affiliation: Beijing National Laboratory for Molecular Sciences (BNLMS), Key Laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China. gqyang@iccas.ac.cn.
authors.forename: Guoqiang
authors.initials: G
authors.lastname: Yang
authors.affiliation: Department of Applied Biology and Chemical Technology and the State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China. kwok-yin.wong@polyu.edu.hk.
authors.forename: Kwok-Yin
authors.identifier: 0000-0003-4984-7109
authors.initials: KY
authors.lastname: Wong
mesh_terms: D015374:Biosensing Techniques; D005456:Fluorescent Dyes; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D025941:Protein Interaction Mapping / Q000379:methods; D013050:Spectrometry, Fluorescence; D013267:Stilbenes; D001618:beta-Lactamases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D013267:Stilbenes; C000617116:tetraphenylethylene; D001618:beta-Lactamases; C034413:beta-lactamase TEM-1