Generic Assays for Functional Characterization (548-591)

膜蛋白在几乎所有的生命过程中都扮演着重要的角色。膜蛋白调节细胞 通过充当信号转导、营养运输、防御外部攻击的门的过程，以及 维持膜动态平衡。然而，由于处理膜蛋白和 对于脂质双层环境的要求，功能研究从来不是直截了当的，需要大量的 努力进行劳动密集型筛选，寻找适合功能鉴定的条件。加速 膜蛋白的功能研究，我们建议开发一个通用的技术研究资源 膜蛋白的功能表征。以我们近十年的研究经验为基础 膜蛋白和我们对十多种新型膜转运蛋白的功能表征， 在通道和酶方面，我们建议在三个子项目中开发互补和协同技术。 目标1(TR和D3.1)侧重于开发微型热电泳法技术，用于分析 膜蛋白的相互作用、动力学和动力学。目标2(tr&d3.2)结合互补的金属氧化物- 具有用于记录膜通道的平面类脂双层膜的半导体(CMOS)芯片。目标3 (TR&D3.3)利用荧光和放射性标记来探测膜转运体和 频道。这三项集成技术将是强大的和具有变革性的，以表征具有挑战性的 膜蛋白。

Membrane proteins play critical roles in virtually all life processes. Membrane proteins regulate cellular processes by functioning as gates for signal transduction, nutrition transport, defense from outside attack, and maintenance of membrane homeostasis. However, due to difficulties in handling membrane proteins and the requirement of a lipid-bilayer environment, functional studies are never straightforward and require substantial efforts in labor-intensive screening for conditions suitable for functional characterization. To accelerate functional studies of membrane proteins, we propose to develop a technical research resource for generic functional characterization of membrane proteins. Built on our nearly ten years of research experiences on membrane proteins and our functional characterization of more than a dozen novel membrane transporters, channels and enzymes, we propose to develop complementary and synergic techniques in three sub-projects. Aim 1 (TR&D3.1) focuses on the development of microscale thermophoresis technology for analysis of the interactions, dynamics and kinetics of membrane proteins. Aim 2 (TR&D3.2) combines the complementary metaloxide- semiconductor (CMOS) chips with the planar lipid bilayer membranes for recording membrane channels. Aim 3 (TR&D3.3) utilizes fluorescent and radioactive labels for probing uptake activities for membrane transporters and channels. The three integrated technologies will be robust and transformative for characterization of challenging membrane proteins.