Enhanced synthetic luciferin substrates for firefly luciferase

萤火虫荧光素酶的增强型合成荧光素底物

• 批准号: 8532676
• 负责人: STEPHEN C. MILLER
• 金额: $ 34.9万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-26 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8532676
• 关键词: Active Biological Transport; Adopted; Affinity; Animal Model; Binding; Biological; Biological Assay; Bioluminescence; Carbon Dioxide; Cells; Chemicals; Cyclization; Cytolysis; Detection; Disease Progression; Drug Efflux; Evaluation; Exhibits; Firefly Luciferases; Fluorescence; Gene Expression; Gene Expression Profiling; Image; In Vitro; Life; Light; Light Cell; Luciferases; Mammalian Cell; Methods; Mutation; Optical reporter; Organism; Output; Penetration; Permeability; Pharmaceutical Preparations; Photochemistry; Property; Pump; Reaction; Reporter Genes; Research; Rotation; Therapeutic; Tissues; Whole Organism; Work; base; bioluminescence imaging; cost; drug candidate; drug discovery; high throughput screening; imaging modality; improved; in vivo; inhibitor/antagonist; light emission; luciferin; mutant; novel therapeutics; protein protein interaction; quantum; response; screening; small molecule

Bioluminescence assays and imaging methods have been widely adopted for drug discovery because of their simplicity, robustness, and low cost. The inherently low background and lack of need for excitation light makes bioluminescence superior to fluorescence methods for many applications. Nevertheless, luciferase has yet to reach its full potential. Light emission from luciferase is limited by 1) access to the luciferin substrate (modulated by affinity, cell-permeability, and active transport by drug pumps), and 2) the photophysical properties of the luciferin (efficiency and wavelength of emission). To maximize the power of luciferase for biological applications we have synthesized new luciferin substrates that enhance luciferase light emission. We propose the following specific aims: 1) synthesis of new luciferin substrates with enhanced light emission properties; 2) identification of mutant luciferases that efficiently utilize synthetic luciferins; 3) evaluation of synthetic luciferins and mutant luciferases for live cell bioluminescence imaging.

生物发光测定和成像方法已被广泛采用， 药物发现，因为它们的简单性，鲁棒性和低成本。固有的低 背景技术和不需要激发光使得生物发光上级 荧光方法用于许多应用。然而，荧光素酶还没有达到 它的全部潜力。来自荧光素酶的光发射受到以下限制：1）接近荧光素酶， 底物（由亲和力、细胞渗透性和药物泵主动转运调节）， 以及2）光致发光的光物理性质（光致发光的效率和波长）。 排放）。为了最大限度地发挥荧光素酶的生物学应用能力， 合成了新的增强荧光素酶发光的荧光素底物。我们提出 具体目标如下：1）合成具有增强的光的新的荧光素底物 发射特性; 2）鉴定有效利用合成的 3）评价用于活细胞的合成的抗肿瘤蛋白和突变的抗肿瘤酶 生物发光成像