Live-Cell Assays of ER and Golgi Enzyme Activities

ER 和高尔基体酶活性的活细胞测定

基本信息

批准号：
8591789
负责人：
JOHN J NALEWAY
金额：
$ 14.98万
依托单位：
MARKER GENE TECHNOLOGIES
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8591789
关键词：
Ally Alzheimer&apos s Disease Amyotrophic Lateral Sclerosis Area Ataxia Biological Biological Assay Biological Monitoring Biotechnology Cell Culture System Cell Culture Techniques Cells Centrifugation Chemicals Coculture Techniques Collaborations Creutzfeldt-Jakob Syndrome Cytolysis Data Detection Development Diagnosis Disease Endoplasmic Reticulum Enzyme Induction Enzyme-Linked Immunosorbent Assay Enzymes Evaluation Exhibits Fluorescence Microscopy Fluorescence Spectroscopy Fluorogenic Substrate Genes Glioblastoma Glycoside Hydrolases Goals Golgi Apparatus High Pressure Liquid Chromatography Huntington Disease In Vitro Individual Intervention Laboratories Lead Life Mannosidase Measurement Measures Medical Medicine Methodology Methods Modeling Multiple System Atrophy Myeloid Leukemia NMR Spectroscopy Neurodegenerative Disorders Non-Insulin-Dependent Diabetes Mellitus Normal Cell Oculocerebrorenal Syndrome Organelles Parkinson Disease Peptide Hydrolases Peptides Permeability Pharmaceutical Preparations Pharmacology Phase Phospholipids Phosphoric Monoester Hydrolases Proteins Protocols documentation Radiolabeled Reproducibility Research Running Sampling Side Small Business Innovation Research Grant Staining method Stains System Techniques Technology Testing Therapeutic Thin Layer Chromatography Time Tissues analytical tool base chemical standard drug discovery drug efficacy efficacy testing enzyme activity enzyme substrate fluorophore in vitro Assay in vitro activity in vivo inhibitor/antagonist innovation meetings novel public health relevance radiotracer research study screening secretase success tissue culture uptake

项目摘要

DESCRIPTION: This Small Business Innovation Research Phase I project aims to develop new targeted fluorogenic substrates capable of measuring enzyme activities in the Golgi apparatus and Endoplasmic Reticulum (ER) of living cells and tissues. If successful, the proposed research will provide breakthroughs needed to advance the discovery of promising new therapies and modulating drugs for neurodegenerative disorders including Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD), Type 2 diabetes, Lowe syndrome, Huntington's disease and allied medical conditions. In Phase I of this project, Marker Gene Technologies, Inc. will establish the feasibility of the technology by preparing new fluorogenic glycosidase, phosphatase and peptidase substrates for enzymes with localized activity in the Golgi and ER and demonstrating differential staining in living cells that are from normal or are of disease origin. In Phase II, these and additional new substrates will be assayed in vitro and in vivo for their ability to measure specific and localized inhibition or induction of these enzymes in living cells as well as differentiate individual enzyme activities in a cell- or tissue-specific manner. The new substrates and the resulting detection systems will provide innovative methods to quantitate Golgi and ER organelle enzyme function and to screen for the influence of secondary drug or protein administration, making them useful analytical tools for drug discovery and diagnosis in a variety of significant medical applications.

描述：这个小型企业创新研究阶段I项目旨在开发能够测量活细胞和组织的高尔基体和内质网（ER）中酶活性的新的靶向荧光底物。如果成功的话，拟议的研究将为您提供有希望的新疗法的突破，并调节包括阿尔茨海默氏病（AD），链肌营养性横向硬化症（ALS），帕金森氏病（PARKINSON），帕克森氏病（PD），2型糖尿病，2型糖尿病，Lowe综合症，匈牙利综合症的药物，以及匈奴的疾病。在该项目的第一阶段，Marker Gene Technologies，Inc。将通过准备新的荧光糖苷酶，磷酸酶和肽酶底物来确定该技术的可行性，以用于在高尔基和ER中具有局部活性的酶，并在正常或疾病起源的活细胞中表现出差异化细胞中的局部活性。在第二阶段，将在体外和体内分析这些和其他新底物，以测量特定和局部抑制或诱导的能力活细胞中的这些酶以及以细胞或组织特异性方式区分单个酶活性。新的底物和结果检测系统将提供创新的方法来定量高尔基和ER细胞器酶功能，并筛选二级药物或蛋白质给药的影响，从而使它们在各种重要的医疗应用中用于药物发现和诊断的有用分析工具。