PEPTIDE/LIPID STUDIES IN FIELD ORIENTED MEMBRANES

描述(改编自申请人的摘要)：尽管有丰富的 明显的膜结合分子的重要性，有戏剧性的缺乏 关于膜系统的高分辨率结构数据。的总体目标 本研究是对新方法论的开发和测试，即 能够提供原子级结构和动态信息 脂双层中的膜结合蛋白。这些方法利用 自发地在磁场中取向的磷脂双层阵列 允许在天然膜的模型中进行高分辨率光谱分析。 这些方法将被应用于跨膜螺旋肽的建模：一 在膜蛋白中发现的基本结构基序。 跨膜螺旋已被证明具有许多重要功能， 包括确定通道和运输路径，定位假体 电子转移反应基团和中介跨膜基团 发信号。 研究的第一步将是优化磁场 用于新型核磁共振研究的可定向脂质双分子膜 跨膜多肽。接下来，具体的顺序和方向 用~2H核磁共振测定模型膜螺旋中的多肽 光谱学。最后，有关 将测量具有周围脂双层的跨膜多肽 同时使用定向2H核磁共振和新型定向电子自旋共振(ESR) 实验。 对原型跨膜螺旋的研究不仅有助于 确定磁定向方法的可行性，但将 我还回答了围绕属性的一些基本问题 跨膜螺旋，以及它们与脂双层的相互作用。 虽然这里提出的研究是针对单个跨膜螺旋的， 这项工作将为研究更大的碎片和完整的 膜蛋白。

DESCRIPTION (Adapted from applicant's abstract): Despite the abundance and clear importance of membrane-bound molecules, there is a dramatic lack of high resolution structural data on membrane systems. The overall goals of this research are the development and testing of new methodology that is capable of providing atomic level structural and dynamic information on membrane-bound proteins in a lipid bilayer. The methods utilize phospholipid bilayer arrays that spontaneously orient in magnetic fields and allow for high resolution spectroscopy in a model for a natural membrane. These methods will be applied to model transmembrane helical peptides: one of the fundamental structural motifs found in membrane proteins. Transmembrane helices have been shown to have many important functions, including defining channels and transport pathways, positioning prosthetic groups for electron transfer reactions, and mediating transmembrane signaling. The first step in the study will be to optimize the magnetic field orientable lipid bilayers for novel nuclear magnetic resonance (NMR) studies of transmembrane peptides. Next, the order and orientation of specific peptides in a model membrane helix will be determined using 2H NMR spectroscopy. Finally, information about the interaction of the transmembrane peptides with the surrounding lipid bilayer will be measured using both oriented 2H NMR and novel oriented electron spin resonance (ESR) experiments. Investigations of prototypal transmembrane helices will not only help establish the feasibility of the magnetic orientation methodology, but will also answer some of the basic questions that surround the properties of transmembrane helices, as well as their interactions with lipid bilayers. Although the studies proposed here are for single transmembrane helices, this work will pave the way for studies of larger fragments and intact membrane proteins.