Generic Assays for Functional Characterization (548-591)

膜蛋白在几乎所有的生命过程中起着至关重要的作用。膜蛋白调节细胞

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.