Flash and grab sodium channel auxiliary subunits with photobleaching microscopy and genetically encoded cross-linkers

我们正在踏上进入现代的分子之旅，在活细胞的环境中进行合成有机化学。我们的方法将结合化学、生物学和显微镜领域的最好技术，以了解心脏中控制电子信号的微小“接收器”的组成和特殊功能。每个心脏细胞都含有数百万个这样的小受体，但我们不知道受体的组成(部分)，也不知道受体的不同部分如何相互作用。值得注意的是，我们的化学和生物技术将使我们能够研究携带一种合成氨基酸的受体，当这种合成氨基酸暴露在某种类型的光下时，这种合成氨基酸会变得活跃，并‘抓住’邻近的受体片段，从而使我们能够识别受体的片段。其他实验将看到我们使用高功率的光来计数受体的片段，方法是测量连接到受体不同部分的遗传编码的“荧光团”的光信号。此外，我们完全现代化的实验方法和尖端的研究环境将使我们能够为我们的人员提供高质量的培训，同时开发对当前研究计划的成功至关重要的新技术，并将使未来的资金申请更具竞争力。学员将学习各种实验技术，包括显微镜、电生理学和有机化学，每一项都将使他们成为学术和工业领域职位的宝贵资产。此外，埃亨实验室一旦开发出这些技术和化学方法，将在整个生物学领域得到广泛应用，并将为加拿大国内外所有水平的学生、本科生、研究生和博士后研究员提供无数培训和合作机会。

We are embarking on a molecular journey into the modern era of performing synthetic organic chemistry in the environment of the living cell. Our approach will combine the best technologies in the fields of chemistry, biology and microscopy to understand the composition and special function of a tiny 'receptor' in the heart that controls electrical signalling. Each heart cell contains millions of these little receptors, but we do not know the composition (parts) of the receptor nor do we know how the different parts of the receptor interact. Of note, our chemical and biological technologies will allow us to study receptors that carry a synthetic amino acid that when exposed to a certain type of light, this synthetic amino acid will become reactive and 'grab' the neighboring piece of the receptor, thus allowing us to identify the pieces of the receptor. Other experiments will see us use high-powered light to count the pieces of the receptor by the measuring light signals from genetically encoded 'flurorophores' that are attached to the different parts of the receptor. Additionally, our thoroughly modern experimental approaches and cutting edge research environment will allow us to provide High Quality Training of our personnel while developing novel techniques that will be invaluable for the success of current research programs and will make future funding applications significantly more competitive. Trainees will learn a variety of experimental techniques, including microscopy, electrophysiology and organic chemistry, each of which that will make them valuable assets for positions in the academic and industrial settings. Moreover, these technologies and chemical approaches, once developed in the Ahern lab, will have broad applications throughout biology and will provide countless training and collaborative opportunities for students of all level, undergraduate, graduate and post-doctoral fellows throughout Canada and abroad.