TY - JOUR
T1 - A HaloTag-Based Multicolor Fluorogenic Sensor Visualizes and Quantifies Proteome Stress in Live Cells Using Solvatochromic and Molecular Rotor-Based Fluorophores
AU - Liu, Yu
AU - Miao, Kun
AU - Li, Yinghao
AU - Fares, Matthew
AU - Chen, Shuyuan
AU - Zhang, Xin
N1 - Funding Information:
*E-mail: xuz31@psu.edu. ORCID Yu Liu: 0000-0002-0779-1488 Kun Miao: 0000-0001-6567-3650 Xin Zhang: 0000-0001-6686-1645 Funding This work has been supported by the Burroughs Wellcome Fund Career Award at the Scientific Interface (X.Z.), a Paul Berg Early Career Professorship (X.Z.), and a Lloyd and Dottie Huck Early Career Award (X.Z.). Notes The authors declare no competing financial interest.
Publisher Copyright:
Copyright © 2018 American Chemical Society.
PY - 2018/8/7
Y1 - 2018/8/7
N2 - Protein homeostasis, or proteostasis, is essential for cellular fitness and viability. Many environmental factors compromise proteostasis, induce global proteome stress, and cause diseases. The proteome stress sensor is a powerful tool for dissecting the mechanism of cellular stress and finding therapeutics that ameliorate these diseases. In this work, we present a multicolor HaloTag-based sensor (named AgHalo) to visualize and quantify proteome stresses in live cells. The current AgHalo sensor is equipped with three fluorogenic probes that turn on fluorescence when the sensor forms either soluble oligomers or insoluble aggregates upon exposure to stress conditions, both in vitro and in cellulo. In addition, AgHalo probes can be combined with commercially available always-fluorescent HaloTag ligands to enable two-color imaging, allowing for direct visualization of the AgHalo sensor both before and after cells are subjected to stress conditions. Finally, pulse-chase experiments can be performed to discern changes in the cellular proteome in live cells by first forming the AgHalo conjugate and then either applying or removing stress at any desired time point. In summary, the AgHalo sensor can be used to visualize and quantify proteome stress in live cells, a task that is difficult to accomplish using previous always-fluorescent methods. This sensor should be suited to evaluating cellular proteostasis under various exogenous stresses, including chemical toxins, drugs, and environmental factors.
AB - Protein homeostasis, or proteostasis, is essential for cellular fitness and viability. Many environmental factors compromise proteostasis, induce global proteome stress, and cause diseases. The proteome stress sensor is a powerful tool for dissecting the mechanism of cellular stress and finding therapeutics that ameliorate these diseases. In this work, we present a multicolor HaloTag-based sensor (named AgHalo) to visualize and quantify proteome stresses in live cells. The current AgHalo sensor is equipped with three fluorogenic probes that turn on fluorescence when the sensor forms either soluble oligomers or insoluble aggregates upon exposure to stress conditions, both in vitro and in cellulo. In addition, AgHalo probes can be combined with commercially available always-fluorescent HaloTag ligands to enable two-color imaging, allowing for direct visualization of the AgHalo sensor both before and after cells are subjected to stress conditions. Finally, pulse-chase experiments can be performed to discern changes in the cellular proteome in live cells by first forming the AgHalo conjugate and then either applying or removing stress at any desired time point. In summary, the AgHalo sensor can be used to visualize and quantify proteome stress in live cells, a task that is difficult to accomplish using previous always-fluorescent methods. This sensor should be suited to evaluating cellular proteostasis under various exogenous stresses, including chemical toxins, drugs, and environmental factors.
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U2 - 10.1021/acs.biochem.8b00135
DO - 10.1021/acs.biochem.8b00135
M3 - Article
C2 - 29474059
AN - SCOPUS:85046078979
SN - 0006-2960
VL - 57
SP - 4663
EP - 4674
JO - Biochemistry
JF - Biochemistry
IS - 31
ER -