PROTEIN SEQUENCER SYSTEM

本提案旨在资助Procise蛋白测序系统。494型由三个集成的分子组成：I) Procise蛋白质测序仪，ii) 190-00型Procise PTH系统，包括140C型微梯度输送系统和785A型紫外检测器，以及iii)配备Procise控制软件和610A型数据分析软件的Macintosh计算机。Procise系统将取代1987年通过DRR-BRS共享仪器拨款购买的过时的Applied Biosystems 477A。新仪器的设计可独立运行多达四个样品盒组件；因此，可以为每个样品定制运行顺序、样品类型和测序方法。它采用多种类型的墨盒组件，包括印迹和微型墨盒，这是定制的不同的样品：液体，PVDF印迹膜和微量。它具有为特定样品开发新方法所需的多功能性。在Edman化学中取代了一种比三乙胺更稳定的偶联碱n -甲基哌啶。这影响了更高的重复产量和进行高灵敏度序列分析的能力。从反应筒到注射回路的内部体积减少到不到以前型号体积的三分之一，从而提高了序列化学的精度和效率。为了更好的峰值检测和集成，Procise有一个内置的模拟到数字转换器，提供以前系统的四倍分辨率。Procise系统的灵敏度、灵活性和先进技术非常适合当今的研究先决条件。这台仪器对于一群对蛋白质错误折叠有着共同兴趣的高产科学家来说是至关重要的，他们对淀粉样蛋白的形成和阿尔茨海默病的发病机制有着共同的兴趣。

This proposal is to fund a Procise Protein Sequencing System. The Model 494 is composed of three integrated molecules: I) the Procise Protein Sequencer, ii) the Model 190-00 Procise PTH System, that includes the Model 140C Microgradient Delivery System and the Model 785A UV Detector, and iii) a Macintosh computer equipped with Procise control software and the Model 610A Data Analysis software. The Procise system will replace an obsolete Applied Biosystems 477A that was purchased in 1987 through a DRR-BRS Shared Instrumentation Grant. The new instrument has been designed to run up to four sample cartridge assemblies independently; thus, the run order, sample type and sequencing method may be customized for each sample. It utilizes multiple types of cartridge assemblies, including the Blot and Micro-cartridge, which are customized for diverse samples: liquid, PVDF blot membranes and micro-quantities. It has the versatility needed to develop new methods for particular samples. A new coupling base, N-methylpiperidine, which is more stable then triethylamine has been substituted in the Edman chemistry. This has effected higher repetitive yields and the ability to perform high- sensitivity sequence analysis. The internal volume from the reaction cartridge to the injection loop has been reduced to less than one-third of the volume in previous models resulting in improved precision and efficiency of the sequence chemistry. For better peak detection and integration, the Procise has a build-in analog to digital converter providing four times the resolution of previous systems. The sensitivity, flexibility and advanced technology of the Procise system is eminently suitable for today's research prerequisites. This instrument would be critical to the needs of a group of highly productive and strongly NIH funded scientists who share a common interest in the misfolding of proteins, with specific focus in amyloid formation and the pathogenesis of Alzheimer's disease.