4 years ago

Visualization of Endoplasmic Reticulum Aminopeptidase 1 under Different Redox Conditions with a Two-Photon Fluorescent Probe

Visualization of Endoplasmic Reticulum Aminopeptidase 1 under Different Redox Conditions with a Two-Photon Fluorescent Probe
Lin Yuan, Weihong Tan, Yong-Chao Liu, Hong-Wen Liu, Jing Zhang, Shuai Xu, Xiao-Bing Zhang, Xiao-Xiao Hu, Feng-Li Qu, Shuang-Yan Huan
Endoplasmic reticulum aminopeptidase 1 (ERAP1), a metallopeptidase belonging to the M1 peptidase family, plays an important role in antigen processing in vivo. Additionally, many diseases are caused by ERAP1 perturbation. Thus, an efficient method for monitoring its content is extremely important for disease diagnosis and treatment. However, few fluorescent probes have been reported for efficiently monitoring ERAP1 in living cells and tissues. In this work, a two-photon fluorescent probe (SNCL) containing 1,8-naphthalimide (two-photon fluorophore), l-leucine (trigger moiety), and a methyl sulfonamide moiety (endoplasmic reticulum-targeting group) for imaging ERAP1 activity in living cells is reported for the first time. The optimized probe exhibited high sensitivity toward ERAP1, with about a 95-fold fluorescence enhancement at 550 nm. Herein, we monitored ERAP1 with SNCL by introducing interferon-γ to induce ERAP1 activity in living cells. The content of ERAP1 was dependent on the redox state of the endoplasmic reticulum, which was demonstrated by using SNCL to monitor the enzymatic activity of ERAP1 under different redox conditions. Excitingly, SNCL was also successfully applied for monitoring ERAP1 in tumor tissue with an imaging depth of 50–120 μm. In conclusion, SNCL not only can be used for the sensitive detection of endogenous ERAP1 in living cells and tumor tissues but also can serve as a potentially useful tool to reveal ERAP1-related diseases.

Publisher URL: http://dx.doi.org/10.1021/acs.analchem.7b01561

DOI: 10.1021/acs.analchem.7b01561

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