Live-Cell Assays of ER and Golgi Enzyme Activities

描述：这个小企业创新研究第一阶段项目旨在开发新的靶向荧光底物，能够测量活细胞和组织的高尔基体和内质网（ER）中的酶活性。如果成功，该研究将提供必要的突破，以推进有希望的神经退行性疾病的新疗法和调节药物的发现，包括阿尔茨海默病（AD）、肌萎缩侧索硬化症（ALS）、帕金森病（PD）、2型糖尿病、洛氏综合征、亨廷顿氏病和相关的医疗条件。在该项目的第一阶段，Marker Gene Technologies, Inc.将通过制备新的荧光糖苷酶、磷酸酶和肽酶底物来确定该技术的可行性，这些底物用于高尔基体和内质网中具有局部活性的酶，并在正常或疾病起源的活细胞中证明差异染色。在第二阶段，这些和额外的新底物将在体外和体内测试它们测量特异性和局部抑制或诱导的能力

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.