HTS driven by fluorescence lifetime detection of FRET identifies activators and inhibitors of cardiac myosin

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

SLAS Discovery Pub Date : 2023-07-01 DOI:10.1016/j.slasd.2023.06.001

JM Muretta , D Rajasekaran , Y Blat , S Little , M Myers , C Nair , B Burdekin , SL Yuen , N Jimenez , P Guhathakurta , A Wilson , AR Thompson , N Surti , D Connors , P Chase , D Harden , CM Barbieri , L Adam , DD Thomas

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引用次数: 3

Abstract

Small molecules that bind to allosteric sites on target proteins to alter protein function are highly sought in drug discovery. High-throughput screening (HTS) assays are needed to facilitate the direct discovery of allosterically active compounds. We have developed technology for high-throughput time-resolved fluorescence lifetime detection of fluorescence resonance energy transfer (FRET), which enables the detection of allosteric modulators by monitoring changes in protein structure. We tested this approach at the industrial scale by adapting an allosteric FRET sensor of cardiac myosin to high-throughput screening (HTS), based on technology provided by Photonic Pharma and the University of Minnesota, and then used the sensor to screen 1.6 million compounds in the HTS facility at Bristol Myers Squibb. The results identified allosteric activators and inhibitors of cardiac myosin that do not compete with ATP binding, demonstrating high potential for FLT-based drug discovery.

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FRET荧光寿命检测驱动的HTS识别心肌肌球蛋白的激活剂和抑制剂

在药物发现中，与靶蛋白上的变构位点结合以改变蛋白质功能的小分子受到高度重视。需要高通量筛选（HTS）分析来促进变构活性化合物的直接发现。我们开发了荧光共振能量转移（FRET）的高通量时间分辨荧光寿命检测技术，该技术能够通过监测蛋白质结构的变化来检测变构调节剂。我们基于光子制药公司和明尼苏达大学提供的技术，通过将心肌肌球蛋白的变构FRET传感器应用于高通量筛选（HTS），在工业规模上测试了这种方法，然后使用该传感器在百时美施贵宝的HTS设施中筛选了160万种化合物。该结果确定了不与ATP结合竞争的心肌肌球蛋白变构激活剂和抑制剂，证明了基于FLT的药物发现的高潜力。

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来源期刊

SLAS Discovery Chemistry-Analytical Chemistry

CiteScore

7.00

自引率

3.20%

发文量

审稿时长

39 days

期刊介绍： Advancing Life Sciences R&D: SLAS Discovery reports how scientists develop and utilize novel technologies and/or approaches to provide and characterize chemical and biological tools to understand and treat human disease. SLAS Discovery is a peer-reviewed journal that publishes scientific reports that enable and improve target validation, evaluate current drug discovery technologies, provide novel research tools, and incorporate research approaches that enhance depth of knowledge and drug discovery success. SLAS Discovery emphasizes scientific and technical advances in target identification/validation (including chemical probes, RNA silencing, gene editing technologies); biomarker discovery; assay development; virtual, medium- or high-throughput screening (biochemical and biological, biophysical, phenotypic, toxicological, ADME); lead generation/optimization; chemical biology; and informatics (data analysis, image analysis, statistics, bio- and chemo-informatics). Review articles on target biology, new paradigms in drug discovery and advances in drug discovery technologies. SLAS Discovery is of particular interest to those involved in analytical chemistry, applied microbiology, automation, biochemistry, bioengineering, biomedical optics, biotechnology, bioinformatics, cell biology, DNA science and technology, genetics, information technology, medicinal chemistry, molecular biology, natural products chemistry, organic chemistry, pharmacology, spectroscopy, and toxicology. SLAS Discovery is a member of the Committee on Publication Ethics (COPE) and was published previously (1996-2016) as the Journal of Biomolecular Screening (JBS).