Specialized Engineering Research Equipment: An LC - Ion Trap Mass Spectrometer for Protein, Peptide and Small Molecule Characterization

本申请的目的是获得一个捆绑的高效液相色谱(HPLC)/LCQ离子阱质谱仪，用于几个研究项目，并用于培训学生。新的LQC离子阱质谱仪能够进行氨基酸测序实验，DNA测序实验，肽磷酸化位点定位和寡糖分析，除了更传统的小分子表征。该设备的使用有以下具体目的：(1)为本科生和研究生提供核心知识和培训，使他们能够在至少三种不同的生物工程应用中使用LCQ离子阱质谱，具体目标2至4中有详细说明；(2)通过蛋白质组学策略对大肠杆菌基因产物进行测序和表征，以增强其蛋白质分泌，并使这些信息在网络上可访问；(3)更好地定义和理解昆虫细胞杆状病毒表达系统中蛋白质糖基化与环境条件的关系，实现模型蛋白的复杂糖基化；(4)表征聚乙二醇（PEG）化的肽和蛋白质，并测试有关大分子药物结构和生物分布的假设。

The objective of this request is to acquire a bundled High Performance Liquid Chromatography (HPLC)/LCQ ion trap mass spectrometer for use in several research projects, and for the training of students. The new LQC ion trap mass spectrometers are able to perform amino acid sequencing experiments, DNA sequencing experiments, mapping of peptide phosphorylation sites and oligosaccharide analysis, in addition to more traditional small molecule characterization. The use of this equipment has the following specific aims: (1) to provide core knowledge and training to undergraduate and graduate students on the use of LCQ ion trap mass spectrometry in at least three different bioengineering applications which are detailed in the specific aims 2 to 4; (2) to sequence and characterize gene products for engineering Escherichia coli with enhanced protein secretion via a proteome science strategy and to make this information web-accessible; (3) to better define and understand the relationship between protein glycosylation and environmental conditions in the insect cell baculovirus expression system to achieve complex glycosylation of a model protein; and (4) to characterize polyethylene glycol (PEG)ylated peptides and proteins, and test hypotheses regarding structure and biodistribution of macromolecular drugs.