Biophysical and molecular characterisation of ORAI channel properties in skin-derived human cell lines

储存操作钙进入（SOCE）的机制在2006年首次被发现是由蛋白STIM和Orai促进的。这种机制现在被认为是控制多种组织功能的关键，特别是它在调节免疫和中枢神经系统中的作用。最近，STIM和Orai已被证明在皮肤角质形成细胞和各种皮肤附属物细胞中介导细胞内Ca2+信号。本提案的目的是使用多学科方法来表征在皮肤来源细胞系中表达的Orai通道同种异构体的生物物理和分子行为。方法将包括细胞培养，膜片钳电生理学和生物信息学/计算生物学。这项工作将在三个地点进行：利物浦大学综合生物学研究所、哈特里中心、STFC达斯伯里和联合利华研究与开发，阳光港。该项目的目的是提供第一个详细的生物物理特征的动力学行为和在皮肤细胞行为背景下的Orai通道的调节潜力。该项目将允许学生：1；推导出他们自己的生物学问题，并通过实验和最先进的计算机模拟来探索这些问题。掌握尖端的膜片钳电生理技术来测量特定的离子通道活性。了解学术的本质：工业合作和工业相关的研究问题

The mechanism of store operated calcium entry (SOCE) in non-excitable tissues was first shown to be facilitated by the proteins STIM and Orai in 2006. This mechanism is now known to be key to controlling a variety of functions in a wide range of tissues, and especially for its role in regulating the immune and central nervous systems. More recently, STIM and Orai have been shown to mediate intracellular Ca2+ signalling in skin keratinocytes and cells from a variety of skin appendages. The objective of this proposal is to use a multi-disciplinary approach to characterise the biophysical and molecular behaviour of Orai channel isoforms expressed in a selection of skin-derived cell lines. Methodologies will include cell culture, patch clamp electrophysiology, and bioinformatics / computational biology. The work will occur at three sites: the Institute of Integrative Biology, University of Liverpool, the Hartree Centre, STFC Daresbury, and Unilever Research & Development, Port Sunlight. The objective of the project is to provide the first detailed biophysical characterisation of the kinetic behaviour and modulatory potential of the Orai channel in the context of skin cell behaviour. The project will allow the student to:1. Derive their own biological questions, and explore these both experimentally and via state-of-the art in silico computer modelling2. Master cutting-edge patch clamp electrophysiology techniques for measuring specific ion channel activities3. Understand the nature of academic:industrial collaborations and industrially-relevant research questions