TY - JOUR
T1 - Light up protein-protein interaction through bioorthogonal incorporation of a turn-on fluorescent probe into β-lactamase
AU - Hu, Rui
AU - Yap, Hong Kin
AU - Fung, Yik Hong
AU - Wang, Yong
AU - Cheong, Wing Lam
AU - So, Lok Yan
AU - Tsang, Chui Shan
AU - Lee, Lawrence Yoon Suk
AU - Lo, Warrick Ken Cheung
AU - Yuan, Jian
AU - Sun, Ning
AU - Leung, Yun Chung
AU - Yang, Guoqiang
AU - Wong, Kwok Yin
N1 - Publisher Copyright:
© The Royal Society of Chemistry.
PY - 2016
Y1 - 2016
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84997047870&partnerID=8YFLogxK
U2 - 10.1039/c6mb00566g
DO - 10.1039/c6mb00566g
M3 - Article
C2 - 27722717
AN - SCOPUS:84997047870
SN - 1742-206X
VL - 12
SP - 3544
EP - 3549
JO - Molecular BioSystems
JF - Molecular BioSystems
IS - 12
ER -