AMINO ACID ANALYZER AND GAS PHASE MICRO SEQUENCER SYSTEM

一组主要调查员代表9个国家要求提供资金。 NIH资助芝加哥洛约拉大学的研究赠款， 一种氨基酸分析仪器系统及其微序列测定方法， 肽和蛋白质的测定。 该系统（包括一个 贝克曼氨基酸分析仪和应用生物系统蛋白质测序仪， 用于高效液相色谱的Perkin-Elmer组件， 惠普小型计算机数据单元）将提供完整的和自动化的 肽和蛋白质一级结构的亚纳摩尔分析 水平 气相微测序仪代表了一个重要的进步， 用于自动Edman降解的液相或固相仪器， 在灵敏度、可靠性和重复产量方面， 被编程以执行偶联、裂解和转化操作， 得到氨基酸苯硫代乙内酰脲衍生物， 鉴定和定量。 HPLC系统提供准确的 溶剂输送和配比系统以及灵敏、稳定的 用于氨基酸苯硫脲分离的可变波长检测器 和侦查 最后，数据单元提供计算机辅助分析 其包括基线减法、吸光度读数的增强， 峰值积分 所要求的氨基酸分析仪和微测序系统代表了 共享的多用户检测包，它将提供 为蛋白质测序提供了一个机构化的设备， 蛋白质结构分析设备 所要求的地点 在这种研究环境中的仪器将加强共享 关于分析和顺序方法的信息，并将促进 名单所列调查人员之间的合作。 该系统的使用将是 由我们的咨询委员会监督（由五名用户加上一名 外部成员），这些成员将对以下方面的职责进行界定： 仪器操作和用户优先级的设置。 研究 微测序仪系统的应用跨越了原核生物和 真核生物分子生物学，膜蛋白和脂蛋白，肽 激素和激素受体，以及各种酶和结构 具有重要生物化学和生理功能的蛋白质。 的 所要求的仪器将在允许序列中发挥关键作用 在每种情况下，在亚纳摩尔水平上进行测定。

Funds are requested by a group of principal investigators representing nine NIH funded research grants at The Loyola University of Chicago to purchase an instrument system for amino acid analysis and the microsequence determination of peptides and proteins. The system (which includes a Beckman Amino Acid Analyzer and an Applied Biosystems Protein Sequencer, Perkin-Elmer components for high performance liquid chromatography and Hewlett-Packard minicomputer data unit) will provide complete and automated analysis of peptide and protein primary structure at the subnanomole level. The gas-phase microsequencer represents an important advance over liquid or solid-phase instrumentation for automated Edman degradation in terms of sensitivity, reliability and repetitive yield; it is flexibly programmed to perform coupling, cleavage and conversion operations, resulting in amino acid phenylthiohydantoin derivatives ready for direct identification and quantitation. The HPLC system provides an accurate solvent delivery and proportioning system as well as a sensitive and stable variable wavelength detector for amino acid phenylthiohydantoin separation and detection. Finally, the data unit provides computer-assisted analysis which includes baseline subtraction, enhancement of absorbance readings and peak integration. The requested amino acid analyzer and microsequencing system represents a shared multiuser instrumentation package which would provide the institutional armamentarium for protein sequence and would provide a protein structural analysis facility. Location of the requested instrumentation within this research environment would enhance the sharing of information on analytical and sequence approaches and would foster collaboration among the listed investigators. Use of the system would be overseen by our Advisory Committee (consisting of five users plus an outside member) which would have defined responsibilities with regard to instrument operation and the setting of user priorities. Research applications of the microsequencer system span the areas of prokaryotic and eukaryotic molecular biology, membrane proteins and lipoproteins, peptide hormones and hormone receptors, and a variety of enzymes and structural proteins with important biochemical and physiological function. The requested instrumentation would play a pivotal role in permitting sequence determination in each of these cases at the subnanomole level.