Nitrooxidoreductase Rv2466c-Dependent Fluorescent Probe for Mycobacterium tuberculosis Diagnosis and Drug Susceptibility Testing
Nitrooxidoreductase Rv2466c-Dependent Fluorescent Probe for Mycobacterium tuberculosis Diagnosis and Drug Susceptibility Testing
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用于结核分枝杆菌诊断和药敏测试的硝基氧化还原酶 Rv2466c 依赖性荧光探针
DOI：
10.1021/acsinfecdis.9b00006
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发表时间：
2019
影响因子：
5.3
通讯作者：
Liu Gang
中科院分区：
文献类型：
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作者：
Mu Ran;Kong Chengcheng;Yu Wenjun;Wang Hongyao;Ma Yao;Li Xueyuan;Wu Jie;Somersan Karakaya Selin;Li Haitao;Sun Zhaogang;Liu Gang
首先，本研究表明，天然产物激发的基于香豆素的硝基呋喃基calanolides（NFCs）可以通过NFC与Rv 2466 c的W21，N51和Y 61结合形成Rv 2466 c-真菌硫醇（MSH）-NFC（RvMN）三元复合物，并被Rv 2466 c特异性还原，这伴随着高水平荧光的产生。此外，结果揭示了MSH的乙酰化半胱氨酸-葡萄糖胺（AcCys-GlcN）基序足以与Rv 2466 c相互作用，并采用完全还原NFC所必需的活性构象。本文进一步的临床转化研究表明，新型荧光NFC探针可以作为一种急需的高通量、低成本的检测方法，用于检测活结核分枝杆菌（Mtb），并可以精确地测定所有可用药物的MIC值。这种方法可以极大地促进表型药物敏感性测试（pDST）的发展，这将允许在诊断后一周内对结核病（TB）进行即时治疗。
Firstly, this study demonstrated that natural product-inspired coumarin-based nitrofuranyl calanolides (NFCs) can form the Rv2466c-mycothiol (MSH)-NFC (RvMN) ternary complex via NFC binding to W21, N51, and Y61 of Rv2466c and be specifically reduced by Rv2466c, which is accompanied by the generation of a high level of fluorescence. Additionally, the results unveiled that the acetylated cysteine–glucosamine (AcCys-GlcN) motif of MSH is sufficient to interact with Rv2466c and adopt the active conformation that is essential for fully reducing NFCs. Further clinical translational investigation in this Article indicated that the novel fluorescent NFC probe can serve as a much needed high-throughput and low-cost detection method for detection of livingMycobacterium tuberculosis(Mtb) and can precisely determine MIC values for a full range of available drugs. This method can greatly facilitate the development of phenotypic drug-susceptibility testing (pDST) that will allow the point-of-care treatment of tuberculosis (TB) within a week after diagnosis.