Development of an ICMT Supported Membrane Sensor

该项目的长期目标是开发基于薄膜的多元传感器阵列。 用于对作用于特定药物的候选治疗剂进行高通量、并行检测的单芯片 膜蛋白靶标。这种传感技术的成功开发可能会加速药物的研发 针对涉及多种疾病的膜蛋白的发现。我们将开发一种稳定的 含异丙基半胱氨酸羧甲基转移酶(ICMT)的不对称膜结构 项目作为癌症化疗药物发现的潜在新工具。ICMT是一种膜蛋白 负责包括RAS在内的-CAAX基序蛋白的羧甲基化的内质网 信号转导蛋白。这种膜传感器结构将使ICMT介导的检测成为可能 模型底物N-乙酰法尼基半胱氨酸的甲基化作为荧光发射的变化 二硫键连接的分子信标的耦合裂解。传感器是从这些不对称的 结构将直接指示候选药物抑制ICMT催化的甲基化的能力。 这一方法将成为筛选药物库中可能存在的先导化合物的有力工具 抑制细胞致癌RAS蛋白的甲基化。这些化合物的发现和发展 是重要的，因为抑制RAS羧甲基化不仅促进RAS的错误定位 蛋白质，但也抑制RAS转化细胞的能力。ICMT是一个优秀的模型系统 这种膜基传感器的开发是因为存在许多表征良好的衬底来提供 数据验证。这些底物将被用作开发高通量筛选方法的工具 这可能会导致对难治性肿瘤的化疗药物的改进。该项目的后续阶段 将讨论多元素传感器阵列的设计、制造、表征和验证 光学透明的衬底。将使用多学科团队方法，结合以下方面的专业知识 生物化学、材料合成和表征、分析化学和实现目标的理论 支撑膜装置。这种探测器阵列概念的未来扩展可能具有深远的影响 有可能加速发现针对许多其他类别的癌症的新的治疗药物 膜相关蛋白。

The long term goal of this project is the development of multi-element membrane-based sensor arrays on a single chip for high-throughput, parallel sensing of therapeutic agent candidates acting on specific membrane protein targets. Successful development of this sensing technology can lead to accelerated drug discovery targeted to membrane proteins involved in a variety of diseases. We will develop a stabilized asymmetric membrane structure containing isoprenylcysteine carboxylmethyltransferase (ICMT) in this project as a potential new tool for drug discovery in cancer chemotherapy. ICMT is a membrane protein in the endoplasmic reticulum responsible for the carboxylmethylation of -CaaX motif proteins, including the Ras signal transduction proteins. This membrane sensor architecture will enable the detection of Icmt-mediated methylation of the model substrate N-acetylfarnesylcysteine as a change in fluorescence emission due to the coupled cleavage of a disulfide-linked molecular beacon. Sensors developed from these asymmetric structures will provide a direct indication of a drug candidate's ability to inhibit methylation catalyzed by Icmt. This approach will serve as a powerful tool for screening drug libraries for lead compounds that are likely to inhibit the methylation of cellular oncogenic Ras proteins. Discovery and development of these compounds are important because inhibition of Ras carboxylmethylation promotes not only the mislocalization of the Ras proteins, but also inhibits the ability of Ras to transform cells. ICMT is an excellent model system for development of this membrane-based sensor because many well-characterized substrates exist to provide data validation. These substrates will be used as tools to develop a high-throughput screening approach that may lead to improved chemotherapeutic agents for refractory tumors. Subsequent phases of the project will address the design, fabrication, characterization, and validation of multi-element sensor arrays on an optically transparent substrate. A multidisciplinary team approach will be used, combining expertise in biochemistry, materials synthesis and characterization, analytical chemistry, and theory to achieve the target supported membrane device. Future extension of this detector array concept could have far reaching potential for accelerating the discovery of new therapeutic agents targeted to many other classes of membrane-associated proteins.