Engineering of membrane protein chemosensors for therapeutic research

描述(由申请人提供)：细胞需要能够检测和响应周围环境中的刺激，包括内源性代谢物、外源性营养物质和环境刺激物。负责这一过程的蛋白质被称为化学感受器，参与调节涉及痛觉、饮食偏好和新陈代谢控制的各种生物过程。由于化学传感器将外部环境信息传递到细胞内，因此几乎所有的化学传感器都是结构复杂的膜蛋白，它们多次跨越细胞的质膜，如离子通道、转运体和G蛋白偶联受体。然而，由于大多数化学传感器很难表达、纯化和增溶，因此关于它们的结构和功能信息特别难获得。这一第二阶段提案的目标是设计具有更高表面表达和更高溶液稳定性的化学传感器，并绘制这些化学传感器上的配体和药物结合位点。

DESCRIPTION (provided by applicant): Cells require the ability to detect and respond to stimuli in their surroundings, including to endogenous metabolites, exogenous nutrients, and environmental irritants. The proteins that are responsible for this process are called chemosensors and are involved in regulating diverse biological processes involved in pain sensation, dietary preference, and metabolic control. Because chemosensors transmit external environmental information into the cell, nearly all chemosensors are structurally-complex membrane proteins that span the cell's plasma membrane multiple times, such as ion channels, transporters, and G protein-coupled receptors. However, because most chemosensors are difficult to express, purify, and solubilize, structural and functional information about them has been exceptionally difficult to obtain. The goal of this Phase 2 proposal is to engineer chemosensors with increased surface expression and increased stability in solution and to map ligand and drug binding sites on these chemosensors.