PURCHASE OF A HIGH SENSITIVITY AMINO ACID ANALYZER

购买高灵敏度氨基酸分析仪

• 批准号: 3521322
• 负责人: WILLIAM C. MERRICK
• 金额: $ 5.4万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-21 至 1992-07-20
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3521322
• 关键词: aminoacid analyzer; biomedical equipment purchase; computer data analysis; protein sequence

This proposal requests funds for the purchase of a high sensitivity amino acid analyzer as part of a new and highly sophisticated instrumentation core. The instrument will be placed in the newly renovated Molecular Biology Core Laboratory which will be completed in July, 1990. The amino acid analyzer is a necessary component to complete the trio which also includes two instruments on hand already, an Applied Biosystems pulsed- liquid phase protein sequencer and a Milligen peptide synthesizer. With the addition of an amino acid analyzer to the Core Laboratory, it will be: 1, possible to more efficiently order samples for sequencing knowing the general amount of material available; 2, possible to characterize samples that fail to sequence due to a blocked amino terminus or insufficient sample; 3, possible to provide a characterization of peptides synthesized in the Core Laboratory routinely rather than await for a time convenient for Dr. Rosenberry and his staff (note that there, high sensitivity is not necessary); 4, provide amino acid composition data as part of the characterization of proteins, especially those isolated from SDS gels. The five major users display a wide variety of interests, but a central theme is the characterization of proteins. Dr. Merrick wishes to determine the primary amino acid sequence of eukaryotic translation factors by a combination of protein and cDNA sequencing. Subsequent studies would focus on protein active sites and site-directed mutagenesis. Dr. Miller is investigating the pathway of protein degradation in Salmonella by characterization of a number endo- and exopeptidases which recognize small peptides as opposed to complete proteins. In addition to characterization of the proteases at the level of primary sequence, active sites and substrate specificity will be examined. Dr. Wood is studying the enzyme transcarboxylase which is made up of three different subunits in a ratio of 12:12:6. Protein chemistry and cloned genes are being examined for active sites and portions of the molecules necessary for subunit-subunit interactions. In a similar manner, enzymes involved with autotrophic growth with CO or CO2 and H2 and the enzymes involved in polyphosphate metabolism will be characterized. Dr. Rosenberry is trying to determine the unique biochemical features of different acetyl-cholinesterase forms in synapses. By cDNA and protein sequencing, unique subunits will be characterized as will be their glycoinositol phospholipid anchors. Dr. Ikebe is studying the regulation of smooth muscle contraction. His focus is on determination of the phosphorylation sites in myocin light chain kinase and caldesmon and the role of phosphorylation in effecting of these two proteins.

该提案要求提供资金，用于购买高灵敏度氨基 酸分析仪作为一个新的和高度复杂的仪器的一部分 核心 该仪器将被放置在新装修的分子 生物学核心实验室，将于1990年7月完成。 氨基 酸分析仪是完成这三个步骤的必要组成部分， 包括两个仪器，一个应用生物系统脉冲- 液相蛋白质测序仪和Milligen肽合成仪。 与 核心实验室增加一台氨基酸分析仪，将： 1，可以更有效地订购样品进行测序， 可用材料的一般数量; 2，可以表征样品 由于氨基末端被封闭或不充分 样品; 3，可能提供合成肽的表征 在核心实验室例行而不是等待方便的时间 罗森伯里博士和他的工作人员（注意，在那里，高灵敏度不是 必要）; 4、提供氨基酸组成数据作为 蛋白质，特别是从SDS凝胶中分离的蛋白质。 这五个主要用户显示了各种各样的兴趣，但一个中心的 主题是蛋白质的特征。 梅里克医生想确定 真核生物翻译因子的一级氨基酸序列 蛋白质和cDNA测序的组合。 随后的研究将集中在 蛋白质活性位点和定点突变。 米勒医生是 研究沙门氏菌蛋白质降解途径， 许多识别小分子的内肽酶和外肽酶的表征 肽而不是完整的蛋白质。 除了表征 蛋白酶在一级序列、活性位点和 将检查底物特异性。 伍德博士正在研究这种酶 转羧酶由三种不同的亚基组成， 十二点十二分六秒 蛋白质化学和克隆基因正在检查活性 亚单位-亚单位所必需的分子的位点和部分 交互. 以类似的方式，参与自养的酶 生长与CO或CO2和H2和酶参与多磷酸盐 代谢将被表征。 罗森伯里博士正在试图确定 不同形式的乙酰胆碱酯酶的独特的生化特征， 突触 通过cDNA和蛋白质测序， 其特征在于它们的糖肌醇磷脂锚定物。 博士 池部正在研究平滑肌收缩的调节。 他的重点 肌球蛋白轻链磷酸化位点的测定 激酶和钙调素的作用以及磷酸化在这些作用中的作用 两种蛋白质