INHALED ANESTHETIC BINDING INTERACTIONS

该项目的长期目标是：1）了解相互作用 负责吸入麻醉剂的结合， 疏水分子，靶向蛋白质和脂质; 2）识别 这些靶标中有利结合位点的结构基序;和3） 确定吸入麻醉剂结合的结构后果 大分子靶点作为理解功能性的第一步 后果该项目将使用直接的光亲和标记 PI最近开发并验证的方法， 在选择的各种模型中的定量麻醉剂结合数据 简单性和可预测性。除了定义的功能， 麻醉剂结合位点，这些定量结合数据将用于 在动态模拟中设置初始条件， 用于协调本程序中其他方法的结果。 实现短期目标有四个具体目标， 表征吸入性麻醉剂如何以及在何处与模型肽结合 和脂质。具体目标1是确定最小结构 氟烷和其他吸入性有机物的结合选择性要求 从麻醉剂到缩氨酸聚-（L-赖氨酸）的均聚物将被研究 其中存在已知的二级结构特征和可操作的 超二级结构这些均聚物将作为支架， 包括其他氨基酸。具体目标2是确定 麻醉剂在合成螺旋束中的结合特征和定位 已知结构。项目二合成的水溶性螺旋束 将被照片标记以确定氟烷结合位置， 特异性和亲和力。系统性氨基酸取代. 选定的位置将允许腔特征与 麻醉剂结合具体目标3是确定特征， 光化学结合到脂质中的氟烷的位置 不同组成的双层。在这一目标下，该项目将与 与项目四有关的初步放置氟烷和 物理组成和变化的影响。具体目标4将决定 脂质/蛋白质界面对于特异性和 功能相关的麻醉剂结合域。绑定字符和 位置将与选择的平面双层中的功能相关。 膜结合的合成Helicobacter。这一目标强烈地与 项目四.

The long term goals of the project are 1) to understand interactions responsible for the binding of inhaled anesthetics, which are small hydrophobic molecules, to target proteins and lipids; 2) to identify structural motifs for favored binding sites in these targets; and 3) to determine structural consequences of inhaled anesthetic binding to macromolecular targets as a first step towards understand functional consequences. The project will use a direct photoaffinity labeling approach recently developed and validated by the PI to provide quantitative anesthetic binding data in a variety of models chosen for their simplicity and predictability. Aside from defining features of anesthetic binding sites, these quantitative binding data will be used to set initial conditions in dynamic simulations, and will also be essential for reconciling results of other approaches in this program. There are four specific aims to accomplish short term objectives, which characterize how and where inhalational anesthetics bind to model peptides and lipids. Specific aim 1 is to determine the minimum structural requirements for binding selectivity of halothane and other inhalational anesthetics to peptides. Homopolymers of poly-(L-lysine) will be studies where there are known secondary structural features and manipulatable supersecondary structures. These hompolymers will serve as a scaffold for the inclusion of other amino acids. Specific aim 2 is to determine anesthetic binding character and location in synthetic helical bundles of known structure. Water soluble helical bundles synthesized in Project II will be photo labeled to determine halothane binding locations, specificities, and affinities. Systematic amino acid substitutions at selected positions will allow correlation of cavity features with anesthetic binding. Specific aim 3 is to determine the character and location of halothane that is photochemically incorporated into lipid bilayers of varying composition. In this aim the project will interact with Project IV regarding the initial placement of halothane and the effect of physical composition and changes. Specific Aim 4 will determine the importance of lipid/protein interfaces regarding specific and functional relevant anesthetic binding domains. Binding character and location will be correlated with function in planar bilayers in selected membrane-incorporated synthetic helicies. This aim strongly interacts with Project IV.