Development of an ICMT Supported Membrane Sensor

ICMT 支持的薄膜传感器的开发

基本信息

批准号：
7560063
负责人：
DAVID H THOMPSON
金额：
$ 38.14万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-03-01 至 2012-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7560063
关键词：
Active Biological Transport Address Adenosylhomocysteinase Adsorption Affinity Amino Acid Motifs Amino Acids Analytical Chemistry Antibodies Architecture Area Binding Biochemical Reaction Biochemistry Biological Biological Assay Biological Models C-terminal Cell membrane Cells Chemotherapy-Oncologic Procedure Chimeric Proteins Computer Systems Development Confocal Microscopy Coupled Cysteine Data Deposition Detection Development Devices Diffusion Dilution Techniques Dimensions Disease Disulfides Elements Endoplasmic Reticulum Engineering Enzymes Epitopes Equilibrium Event Exposure to Family Film Flowcharts Fluorescence Fluorescence Recovery After Photobleaching Future Generations Goals Green Fluorescent Proteins Hemagglutinin Histidine Homocysteine Homocystine Hydrolase Hydrolysis Immunofluorescence Immunologic Integral Membrane Protein Interference Microscopy Intervention Ionic Strengths Lateral Lead Libraries Link Lipids Liposomes Maintenance Mattresses Measures Mechanics Mediating Membrane Membrane Proteins Methods Methylation Micelles Mission Modeling Molecular Molecular Conformation Molecular Weight Monitor Monte Carlo Method Nitrilotriacetic Acid Nutrient Oncogenic Operative Surgical Procedures Outcome Performance Pharmaceutical Preparations Phase Play Polyethylene Glycols Polyethylenes Post-Translational Protein Processing Preclinical Drug Evaluation Principal Investigator Production Property Proteins Reaction Refractory Reporter Research Research Personnel Role Saccharomyces cerevisiae Scheme Side Signal Transduction Silicon Dioxide Solid Solutions Structure Subarachnoid Hemorrhage Sulfhydryl Compounds Surface System Techniques Technology Temperature Testing Therapeutic Agents Thick Time Validation Variant Vesicle base cell transformation chemotherapeutic agent density design detector drug candidate drug development drug discovery extracellular fluorophore functional group high throughput screening improved inhibitor/antagonist innovation isoprenylcysteine carboxylmethyltransferase member milligram multidisciplinary novel novel strategies novel therapeutics pathogen prevent programs protein expression protein function ras Proteins receptor reconstitution research study response sensor theories tool tumor vapor

项目摘要

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.

该项目的长期目标是开发基于膜的多元件传感器阵列单芯片，用于对作用于特定药物的候选治疗剂进行高通量、并行传感膜蛋白靶标。这种传感技术的成功开发可以加速药物研发针对参与多种疾病的膜蛋白的发现。我们将开发稳定的含有异戊二烯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是一种膜蛋白内质网负责 -CaaX 基序蛋白（包括 Ras）的羧甲基化信号转导蛋白。这种膜传感器架构将能够检测 Icmt 介导的模型底物 N-乙酰法呢基半胱氨酸的甲基化作为荧光发射的变化，由于二硫键连接的分子信标的耦合裂解。由这些不对称发展而来的传感器结构将直接表明候选药物抑制 Icmt 催化的甲基化的能力。这种方法将成为筛选药物库中可能的先导化合物的强大工具。抑制细胞致癌 Ras 蛋白的甲基化。这些化合物的发现和开发很重要，因为抑制 Ras 羧甲基化不仅会促进 Ras 的错误定位蛋白，而且还抑制 Ras 转化细胞的能力。 ICMT 是一个优秀的模型系统开发这种基于膜的传感器是因为存在许多特性良好的基板来提供数据验证。这些底物将用作开发高通量筛选方法的工具这可能会导致难治性肿瘤化疗药物的改进。项目的后续阶段将解决多元件传感器阵列的设计、制造、表征和验证光学透明基材。将采用多学科团队方法，结合各领域的专业知识生物化学、材料合成和表征、分析化学以及实现目标的理论支持膜装置。这种探测器阵列概念的未来扩展可能会产生深远的影响加速发现针对许多其他类别的新治疗剂的潜力膜相关蛋白。