Protein-Membrane Organization and Function: Ion Channels

蛋白质膜组织和功能：离子通道

• 批准号: 9313835
• 负责人: James Hinton
• 金额: $ 24万
• 依托单位: University of Arkansas
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-05-15 至 2000-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9313835&HistoricalAwards=false
• 关键词: Protein; Membrane; Organization; Function; Ion

9313835 Hinton The transport of matter through membranes is a basic phenomenon of life. All organisms are separated from their environment by special membrane barriers which control the exchange of matter with their surroundings and act as an electrical resistor. In addition, nearly all life processes are intimately involved with membrane function. Thus, membrane functions play a central role in communication between and within cells, in the transformation of metabolic energy into osmotic, electrical, and to some extent mechanical work. In living cells an electrical potential difference exists between the cytoplasma and the extracellular medium because of the unequal distribution of ions on both sides of the plasma membrane surrounding the cell. The cell membranes are equipped with special transport mechanisms which accelerate the passage of ions across the lipid barrier. An understanding of these transport mechanisms is a vital link to the ultimate goal of the comprehension of some types of membrane function. The transport of monovalent cations, such as Li+, Na+, and K+, through cell membranes plays a key role in many different physiological processes. These processes involve the maintenance of transmembrane potential, conduction of nerve impulses down the axon and across the neuromuscular junction and the differentiation and growth of cells. There are relatively few physical techniques available for the characterization of the interaction of monovalent cations with biological molecules responsible for cation transport. However, nuclear magnetic resonance spectroscopy (NMR) has proven to be an exceptionally powerful technique for the study of the transport process and the molecular systems responsible for the transport. NMR methods may be used to determine the 3-dimensional structure of the channel, to determine the thermodynamic parameters for the integration of transport system into membranes, to determine the thermodynamic parameters for the binding of the cations to the channel and for determining the activation enthalpy for the transport process. %%% The objectives for the requested funding period involve the use of multinuclei multidimensional NMR spectroscopy, structure determination through the use of NMR determined proton distance restrained molecular dynamics/simulated annealing and peptide synthesis to: (1) study the effect of single amino acid substitution on channel structure, the integration into a membrane as a channel, cation binding and transport in the channel system, gramicidin. The objective is to determine how the structure is related to channel formation and selective monovalent cation transport; (2) to obtain the structure of the model membrane bound individual helical components of the pentameric helical bundle that is related to the nicotinic acetylcholine receptor channel. Monovalent cation binding and transport selectivity will also be investigated. ***

9313835欣顿物质通过膜的运输是生命的基本现象。 所有的生物体都被特殊的膜屏障与它们的环境隔开，这些膜屏障控制着物质与周围环境的交换，并起到电阻器的作用。 此外，几乎所有的生命过程都与膜功能密切相关。 因此，膜功能在细胞之间和细胞内的交流中，在代谢能转化为渗透功、电功和某种程度上的机械功中发挥着核心作用。 在活细胞中，细胞质和细胞外介质之间存在电位差，这是因为围绕细胞的质膜两侧的离子分布不均匀。 细胞膜具有特殊的转运机制，可加速离子穿过脂质屏障。 理解这些转运机制是理解某些类型的膜功能的最终目标的重要环节。 一价阳离子如Li+、Na+和K+通过细胞膜的转运在许多不同的生理过程中起着关键作用。 这些过程涉及跨膜电位的维持、神经冲动沿轴突向下和穿过神经肌肉接头的传导以及细胞的分化和生长。 可用于表征一价阳离子与负责阳离子转运的生物分子相互作用的物理技术相对较少。 然而，核磁共振光谱（NMR）已被证明是一种非常强大的技术，用于研究运输过程和负责运输的分子系统。 NMR方法可用于确定通道的三维结构，确定用于将转运系统整合到膜中的热力学参数，确定用于阳离子与通道结合的热力学参数，以及确定用于转运过程的活化焓。 申请资助期间的目标包括使用多核多维NMR光谱学，通过使用NMR确定的质子距离限制分子动力学/模拟退火和肽合成进行结构测定，以：（1）研究单个氨基酸取代对通道结构的影响，作为通道整合到膜中，阳离子结合和通道系统中的运输，短杆菌肽。 目的是确定结构如何与通道形成和选择性单价阳离子转运相关;（2）获得与烟碱乙酰胆碱受体通道相关的五聚体螺旋束的模型膜结合单个螺旋组分的结构。 单价阳离子结合和运输选择性也将进行研究。 ***