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

我们正踏上一段分子之旅，进入在活细胞环境中进行合成有机化学的现代时代。我们的方法将结合化学、生物学和显微镜领域的最佳技术，以了解心脏中控制电信号的微小“受体”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，但我们既不知道受体的组成（部分），也不知道受体的不同部分是如何相互作用的。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合成氨基酸的受体，当暴露在某种类型的光下时，这种合成氨基酸将变得活跃并“抓住”受体的邻近片段，从而使我们能够识别受体的片段。在其他实验中，我们将使用高倍光，通过测量附着在受体不同部位的基因编码“荧光团”发出的光信号，来计算受体的片段。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在开发新技术的同时为我们的人员提供高质量的培训，这将对当前研究项目的成功具有不可估量的价值，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环境中有价值的资产。此外，这些技术和化学方法，一旦在埃亨实验室开发出来，将在整个生物学领域有广泛的应用，并将为加拿大和国外所有水平的学生，本科生，研究生和博士后提供无数的培训和合作机会。

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.