Enhanced synthetic luciferin substrates for firefly luciferase

生物发光分析和成像方法已被广泛应用于 药物发现，因为它们简单、健壮和低成本。与生俱来的低 背景和不需要激发光使生物发光优于 荧光方法有多种应用。尽管如此，荧光素酶还没有达到 它的全部潜力。荧光素酶的光发射受限于1)对荧光素的访问 底物(受亲和力、细胞渗透性和药物泵主动转运的调节)， 2)荧光素的光物理性质(效率和波长 排放)。为了最大限度地发挥荧光素酶在生物应用中的作用，我们有 合成了增强荧光素酶发光的新的荧光素底物。我们建议 具体目标如下：1)合成新型增强型荧光素底物 发光特性；2)有效利用人工合成的突变的荧光素酶的鉴定 荧光素；3)活细胞合成荧光素和突变的荧光素酶的评价 生物发光成像。

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.