INHALED ANESTHETIC BINDING INTERACTIONS

该项目的长期目标是1)了解互动 负责吸入性麻醉剂的结合，这些麻醉剂很小 疏水分子，靶向蛋白质和脂类；2)鉴定 这些靶标中有利结合位点的结构基序；以及3)至 确定吸入麻醉剂结合的结构性后果 作为了解功能的第一步的大分子靶标 后果。该项目将使用直接的光亲和标记 PI最近开发和验证的方法，以提供 选择的各种模型的麻醉剂定量结合数据 它们的简单性和可预测性。除了定义以下功能之外 ，这些定量结合数据将被用来 在运动仿真中设置初始条件，这也将是必不可少的 用于协调本计划中其他方法的结果。 实现短期目标有四个具体目标，分别是 描述吸入麻醉剂如何以及在哪里与模型多肽结合 还有血脂。具体目标1是确定最小结构 氟烷和其他吸入剂结合选择性的要求 从麻醉剂到多肽。将研究聚(L-赖氨酸)均聚物 其中有已知的次级结构特征和可操纵的 超次级结构。这些均聚体将作为 其他氨基酸的包含。具体目标2是确定 麻醉剂在合成螺旋束中的结合特性和位置 已知的结构。项目二中合成的水溶性螺旋束 将被照片标记以确定氟烷结合位置， 特殊性和亲和力。系统的氨基酸替换在 所选位置将允许将型腔特征与 麻醉剂捆绑。具体目标3是确定角色和 光化学结合到脂类中的氟烷的位置 组成各不相同的双层膜。在这一目标中，该项目将相互作用 与项目IV关于氟烷的初始放置和 身体成分和变化的影响。具体目标4将决定 脂/蛋白界面的重要性与特异性和 功能相关的麻醉剂结合域。绑定字符和 位置将与选定的平面双层中的功能相关联 膜结合的合成螺旋体。这一目标与 项目IV。

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.