Nanostructures for Solubilizing Serpentine Receptors

描述(申请人提供)：膜蛋白代表一种大的 所有细胞蛋白质的比例，在细胞过程中起着关键作用 并与人类疾病的许多方面有关。膜技术中的一个难点 蛋白质研究一般认为，这类蛋白质很难 通过重组形式表达、纯化和表征结构 X射线结晶学或其他技术。问题产生于 维持混合的极地和疏水环境的必要性 蛋白质的结构和功能的完整性。我们发现了一个 用于溶解膜蛋白的纳米磷脂组装体 提供脂质体和洗涤剂胶束的有益方面。这个 这项技术是基于磷脂和膜的双层组装 自组装形成纳米磷脂的支架蛋白(MSP) 双层圆盘，MSP使颗粒稳定在圆周上 双层结构域。将膜蛋白插入双分子层导致 单体增溶物种，可在溶液中或表面进行研究。 特别令人感兴趣的是G-蛋白的增溶和稳定 偶联受体，或蛇纹石受体，是 信号转导。GPCRs是一类重要而多样的受体 它结合了几类具有药理意义的生物活性配体。这个 开发纳米胆层组件的实验策略包括 旨在最小化MSP结构的分子工程和 通过以下方式提高纳米膜实体的稳定性和单分散性 改变母体分子的结构。MSP是修改后的形式 人载脂蛋白A-I，循环高密度脂蛋白的一种成分 在胆固醇运输和动脉粥样硬化中发挥作用。我们的发现 对理解脂蛋白的结构也有一定的意义。 MSP-双层结构也将被设计成允许操纵 在固体载体上掺入膜蛋白用于研究 扫描探针显微镜和表面等离子激元的表面敏感技术 共鸣。所开发的材料和技术将在 生物技术和结构/功能生物学研究领域 相关、结构确定、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.