Dynamics and folding of a membrane protein and transcription activator

脂质膜包裹着所有动物、植物和细菌细胞。嵌入脂质中的蛋白质通道和受体精确地控制着营养物质、废物、能量和信息进出细胞的流动，但人们对这些通道和受体的了解甚少。我们的长期目标是开发方法来克服膜蛋白结构，折叠和动力学分析中的许多困难。近年来，我们在研究细菌膜蛋白的折叠，甘油促进剂在洗涤剂环境中取得了进展。我们建议采用一种新的方法，利用细胞提取物合成蛋白质，并研究蛋白质在脂质膜环境中的折叠。无细胞合成开辟了用同位素标记蛋白质以进行核磁共振（NMR）光谱分析的可能性，这是一种类似于磁共振成像（MRI）的方法。通过标记蛋白质，我们将研究蛋白质的详细原子动力学，这是理解其生物运输机制的关键。我们还将沿着类似的路线继续研究真菌抗生素肽alamethicin，这是一种模型膜蛋白。

A lipid membrane surrounds all animal, plant, and bacterial cells. Protein channels and receptors embedded in the lipid precisely control the flow of nutrients, waste products, energy, and information into and out of cells, but are poorly understood. Our long-term goal is to develop methods to overcome the many difficulties in membrane protein structure, folding and dynamics analysis. Recently, we have made progress in studying the folding of a bacterial membrane protein, the glycerol facilitator in detergent environments. We propose to continue this work by applying a new method for synthesizing the protein using cellular extracts and studying folding of the protein in a lipid membrane environment. Cell-free synthesis opens up the possibility of labelling the protein with isotopes for analysis by Nuclear Magnetic Resonance (NMR) spectroscopy, a method analogous to Magnetic Resonance Imaging (MRI). With labeled protein, we will study the detailed atomic dynamics of the protein that are key to understanding its biological transport mechanism. We will also continue our work along similar lines on the fungal antibiotic peptide alamethicin that is a model membrane protein.