TY - JOUR
T1 - Off-On Squalene Epoxidase-Specific Fluorescent Probe for Fast Imaging in Living Cells
AU - Zang, Tienan
AU - Wang, Shudong
AU - Su, Sa
AU - Gao, Mengxu
AU - Chen, Qianqian
AU - Liang, Chenlu
AU - Jing, Jing
AU - Zhang, Rubo
AU - Zhang, Xiaoling
N1 - Publisher Copyright:
© 2021 American Chemical Society
PY - 2021/11/9
Y1 - 2021/11/9
N2 - SQLE (squalene epoxidase) is a cell membrane-bound enzyme. It is a target of fungicides and may become a new target for cancer therapy. Therefore, monitoring the content and distribution of the key enzyme in living cells is very challenging. To achieve this goal, tetraphenyl ethylene-Ter (TPE-Ter) was first designed as a new fluorescent probe to SQLE based on its active cavity. Spectral experiments discovered that SQLE/TPE-Ter shows stronger emission with fast response time and low interference from other analytes. Molecular dynamics simulation clearly confirmed the complex structure of SQLE/TPE-Ter, and the key residues contribute to restriction of TPE-Ter single-molecular motion in the cavity. TPE-Ter-specific response to SQLE is successfully demonstrated in living cells such as LO2, HepG2, and fungi. Imaging of TPE-Ter-treated fungi indicates that it can be used to rapidly assess antifungal drug susceptibility (30 min at least). The present work provides a powerful tool to detect content and distribution of SQLE in living cells.
AB - SQLE (squalene epoxidase) is a cell membrane-bound enzyme. It is a target of fungicides and may become a new target for cancer therapy. Therefore, monitoring the content and distribution of the key enzyme in living cells is very challenging. To achieve this goal, tetraphenyl ethylene-Ter (TPE-Ter) was first designed as a new fluorescent probe to SQLE based on its active cavity. Spectral experiments discovered that SQLE/TPE-Ter shows stronger emission with fast response time and low interference from other analytes. Molecular dynamics simulation clearly confirmed the complex structure of SQLE/TPE-Ter, and the key residues contribute to restriction of TPE-Ter single-molecular motion in the cavity. TPE-Ter-specific response to SQLE is successfully demonstrated in living cells such as LO2, HepG2, and fungi. Imaging of TPE-Ter-treated fungi indicates that it can be used to rapidly assess antifungal drug susceptibility (30 min at least). The present work provides a powerful tool to detect content and distribution of SQLE in living cells.
UR - http://www.scopus.com/inward/record.url?scp=85118782390&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.1c03168
DO - 10.1021/acs.analchem.1c03168
M3 - Article
AN - SCOPUS:85118782390
SN - 0003-2700
VL - 93
SP - 14716
EP - 14721
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 44
ER -
