Endoplasmic Reticulum Associated Degradation (ERAD) of Membrane Proteins in Yeast

总结/摘要 请NIGMS提供资金，用于购买一台阿默舍姆Typhoon IP磷光成像仪，沿着8台 成像板和盒、容纳板的成像托盘以及数据软件许可证 远程计算机分析。反过来，匹兹堡大学和生物科学系 已承诺资金直接抵消成像仪的成本（6，000美元），购买计算机（约2，500美元）， 并提供长期维护合同。成像仪可快速扫描高达34 x 46 cm的凝胶和平板， 产生高分辨率的数字，在长动态范围内产生定量数据，并具有最先进的 软件，以适应不同的应用程序。与此相反，放射性同位素为基础的实验结果， Brodsky和Berman实验室目前依赖于一台使用8年的GE Typhoon弗拉7000磷光成像仪。这个成像仪 使用旧的软件包，该软件包与扫描软件没有链接，用户界面不友好， 机械和软件故障，并产生质量较低的图像，限制了数据分析， 定量相机也只能容纳小于20 x 40 cm的凝胶。购买该 阿默舍姆台风IP磷成像仪与向Jeffrey Brodsky教授授予R 01 GM 075061和资助有关 R 01 GM 116889致Andrea Berman教授。授权R 01 GM 075061中概述的最关键实验， 教授Brodsky要求使用放射性同位素，这允许对选择进行定量评估， 泛素化和内质网中错误折叠蛋白的降解。因此，最佳 一个有效的磷光成像仪对于实现该项目的每一个目标都至关重要。此外，申请 Brodsky教授的MIRA（R35 GM 131732）获得批准，申请开始日期为5月1日， 2019.在最近批准的拨款申请中提出的实验还需要使用 放射性同位素和磷光成像仪来实现项目的每一个目标。共同采购 阿默舍姆台风IP磷光成像仪对教授的近期和长期目标都至关重要。 布罗德斯基的研究项目。

Summary/Abstract Funds from NIGMS are requested for the purchase of an Amersham Typhoon IP phosphorimager along with 8 imaging plates and cassettes, an imaging tray to accommodate the plates, and software licenses for data analysis on remote computers. In turn, the University of Pittsburgh and Department of Biological Sciences have committed funds to directly off-set the cost of the imager ($6,000), to purchase the computer (~$2,500), and to provide a long-term maintenance contract. The imager rapidly scans gels and plates up to 34 x 46 cm, produces high resolution figures, yields quantitative data over a long dynamic range, and has state-of-the-art software to accommodate diverse applications. In contrast, results of radioisotope-based experiments in the Brodsky and Berman labs currently rely on an 8-year old GE Typhoon FLA 7000 phosphorimager. This imager uses an older software package that is unlinked from the scanning software and not user-friendly, suffers from mechanical and software failures, and produces lower quality images that restrict data analysis and quantitation. The imager also only accommodates gels that are smaller than 20 x 40 cm. The purchase of the Amersham Typhoon IP phosphorimager is linked to grant R01 GM075061 to Prof. Jeffrey Brodsky and grant R01 GM116889 to Prof. Andrea Berman. The most critical experiments outlined in grant R01 GM075061 to Prof. Brodsky require the use of radioisotopes, which allows for quantitative assessment of the selection, ubiquitination, and degradation of misfolded proteins in the endoplasmic reticulum. Thus, an optimally functioning phosphorimager is essential to achieve each of the project’s goals. Furthermore, an application for a MIRA (R35 GM131732) to Prof. Brodsky was approved for funding with a requested start date of May 1, 2019. The experiments proposed in this recently approved grant application also require the use of radioisotopes and thus a phosphorimager to achieve each of the project’s goals. Together, the procurement of the Amersham Typhoon IP phosphorimager is critical for both the immediate and longer-term goals of Prof. Brodsky’s research program.