Liquid Chormatograph Mass Spectrometer (LC-MS)

该提案要求提供资金购买一台液相色谱仪(LC-MS)，以支持五个主要用户(全部由美国国立卫生研究院资助)和两个次要用户(一个由美国国立卫生研究院资助)的研究。所有主要和次要用户都是特拉华大学化学和生物化学系的教员。该仪器将放置在一个开放的实验室中，由部门质谱学设施监督。无需预约的用户将能够从几种预先建立的方法中选择一种，这些方法是为快速样品周转而设计的。更复杂的实验将在质谱仪设施人员的协助下进行，并将以最大限度地减少对无预约使用的影响的方式进行安排。拟议的LC-MS仪器将促进几个正在进行的研究项目。这些项目包括：(Colman)鉴定新的亲和标记、化学修饰的酶、纯化的修饰多肽和突变酶；(KOH)选择性调控小分子和光激活基因表达的基因表达，以控制基因表达的时空；(Mueller)RNA修饰、伪尿苷合成酶和4-硫代尿苷生成的酶学；(Riordan)乙酰辅酶A合成酶的模型研究；(Thorpe)脂肪酸代谢中的黄素蛋白、酰基辅酶A脱氢酶和脂肪酸β氧化相关酶以及磺基氧化酶催化。

This proposal requests funds to purchase a liquid chromatograph mass spectrometer (LC-MS) to support the research of five major users (all NIH funded) and two minor users (one NIH funded). All major and minor users are faculty in the chemistry and biochemistry department at the University of Delaware. The instrument will be placed in an open- access laboratory under the supervision of the departmental mass spectrometry facility. Walk-in users will be able to choose one of several pre-established methods that are designed for fast sample turn around. More sophisticated experiments will be performed with assistance from personnel in the mass spectrometry facility and will be scheduled in a manner that minimizes the impact upon walk-in use. The proposed LC-MS instrument will facilitate several ongoing research projects. These projects include: (Colman) characterization of new affinity labels, chemically modified enzymes and purified modified peptides and mutant enzymes; (Koh) selective regulation of gene expression with small molecules and light activated gene expression for temporal and spatial control of gene expression; (Mueller) enzymology of RNA modification, pseudouridine synthases and 4-thiouridine generation; (Riordan) model studies of acetyl coenzyme A synthase; (Thorpe) flavoproteins in fatty acid metabolism, acyl coenzyme A dehydrogenases and related enzymes of fatty acid beta-oxidation, and sulfhydryl oxidase catalysis.