Nanostructures for Solubilizing Serpentine Receptors

描述（由申请人提供）：膜蛋白代表了一种大的 所有细胞蛋白质的比例，在细胞过程中发挥关键作用 并且与人类疾病的许多方面有关。膜的困难 一般来说，蛋白质的研究是这类蛋白质很难 以重组形式表达、纯化和结构表征， X射线晶体学或其它技术。问题来自于 维持混合极性和疏水环境的必要性， 蛋白质的结构和功能完整性。我们已经发现了一种 用于溶解膜蛋白的纳米级磷脂组装体， 提供了脂质体和去污剂胶束的有益方面。的 该技术基于磷脂和膜的双层组件 支架蛋白（MSP）自组装形成纳米级磷脂 具有MSP的双层盘在双层盘的周边稳定颗粒， 双层结构域。将膜蛋白插入到双层中导致 可以在溶液中或表面上研究的单体增溶物质。 特别感兴趣的是G蛋白的溶解和稳定 偶联受体（GPCR）或蛇形受体，它们是 信号转导GPCR是一类重要且多样的受体 其结合几类药理学上感兴趣的生物活性配体。的 开发纳米双层组件的实验策略包括 分子工程旨在最小化MSP的结构， 增加纳米双层实体的稳定性和单分散性， 改变母体分子的结构。MSP是一种修改后的 人载脂蛋白A-I，循环高密度脂蛋白的一种组分 在胆固醇运输和动脉粥样硬化中发挥作用。我们的发现 这对了解脂蛋白的结构也有意义。 MSP-双层结构也将被工程化以允许操纵 在固体支持物上掺入膜蛋白， 扫描探针显微镜和表面等离子体表面敏感技术 共振开发的材料和技术将在以下方面具有潜在的用途： 生物技术和结构/功能生物学研究领域 关联、结构测定、生物分离和药物发现。

DESCRIPTION (provided by applicant): Membrane proteins represent a large proportion of all cellular proteins, play critical roles in cellular processes and are involved in many aspects of human disease. A difficulty in membrane protein research in general is that this class of proteins is difficult to express in recombinant form, purify, and characterize structurally through x-ray crystallography or other techniques. The problem arises from the necessity of maintaining mixed polar and hydrophobic environments to maintain the structural and functional integrity of the protein. We have discovered a nanoscopic phospholipid assembly for solubilizing membrane proteins that provides beneficial aspects of both liposomes and detergent micelles. The technology is based on bilayer assemblies of phospholipid and a membrane scaffold protein (MSP) which self-assemble to form nanoscopic phospholipid bilayer disks with the MSP stabilizing the particle at the perimeter of the bilayer domain. Insertion of membrane proteins into the bilayer results in monomeric solubilized species which can be studied in solution or on surfaces. Of particular interest is the solubilization and stabilization of G-protein coupled receptors (GPCR's), or serpentine receptors, which are components of signal transduction. GPCR's are an important and diverse class of receptors that bind several classes of bioactive ligands of pharmacological interest. The experimental strategy for development of the nanobilayer assembly includes molecular engineering aimed at minimizing the structure of the MSP and increasing the stability and monodispersity of the nanobilayer entity by altering the structure of the parent molecule. The MSP is a modified form of human apolipoprotein A-I, a component of circulatory high-density lipoproteins that play a role in cholesterol transport and atherosclerosis. Our discovery also has implications for understanding the structure of lipoproteins as well. The MSP-bilayer structure will also be engineered to allow manipulation of incorporated membrane proteins on solid supports for study by the surface-sensitive techniques of scanning probe microscopy and surface plasmon resonance. The materials and techniques developed will be potentially useful in areas of biotechnology and biological research for structure/function correlation, structure determination, bioseparation, and drug discovery.