A novel firefly luciferase biosensor enhances the detection of apoptosis induced by ESAT-6 family proteins of Mycobacterium tuberculosis

被引：19

作者：

Shi, Junwei ^{[1
]}

Zhang, Huan ^{[1
]}

Fang, Liurong ^{[1
]}

Xi, Yongqiang ^{[1
]}

Zhou, Yanrong ^{[1
]}

Luo, Rui ^{[1
]}

Wang, Dang ^{[1
]}

Xiao, Shaobo ^{[1
]}

Chen, Huanchun ^{[1
]}

机构：

[1] Huazhong Agr Univ, Coll Vet Med, State Key Lab Agr Microbiol, Wuhan 430070, Peoples R China

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2014年 / 452卷 / 04期

基金：

中国国家自然科学基金;

关键词：

Caspase-3; Firefly luciferase; Apoptosis; Biosensor; Mycobacterium tuberculosis; NF-kappa B; CELL-DEATH; MACROPHAGES; GENES;

D O I：

10.1016/j.bbrc.2014.09.047

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The activation of caspase-3 is a key surrogate marker for detecting apoptosis. To quantitate caspase-3 activity, we constructed a biosensor comprising a recombinant firefly luciferase containing a caspase-3 cleavage site. When apoptosis was induced, caspase-3 cleavage of the biosensor activated firefly luciferase by a factor greater than 25. The assay conveniently detected apoptosis in real time, indicating that it will facilitate drug discovery. We screened ESAT-6 family proteins of Mycobacterium tuberculosis and found that esxA, esxT and esxL induced apoptosis. Further, activation of nuclear factor-kappa B (NF-kappa B) and the NF-kappa B-regulated genes encoding tumor necrosis factor-alpha (TNF-alpha) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) participated in esxT-induced apoptosis. We conclude that this assay is useful for high-throughput screening to identify and characterize proteins and drugs that regulate apoptosis. (C) 2014 Elsevier Inc. All rights reserved.

引用

页码：1046 / 1053

页数：8

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