OLD YELLOW ENZYME--CLONING AND STRUCTURE

我们一直在使用通用临床研究中心维护的GCG包进行老黄酶(OYE)的研究。从酵母菌中克隆了OYE的三种同工酶，并进行了序列测定。这些序列具有很高的同源性，但Oye的生理功能尚不清楚。我们继续寻找具有序列同源性的酶，试图确定其功能。OYE的一个同工酶的晶体结构已经解决，我们一直在突变显着残基，以确定该酶对配体结合和NADPH氧化的影响。我们计划使用GCG程序的几个功能来设计额外突变的引物。该实验室的总体重点是研究含有核黄素的酶。目前，我们正在进行D-氨基酸氧化酶和2-甲基-3-羟基吡啶-5-羧酸加氧酶的克隆和鉴定。在这些研究中，我们将继续使用GCG包的功能。

We have been using the GCG package maintained by the General Clinical Research Center in the study of Old Yellow Enzyme (OYE). Three isozymes of OYE from Saccharomyces have been cloned and sequenced. These sequences have a high homology, but the physiological function of OYE is still unknown. We continue to look for enzymes with sequence homology in an attempt to identify function. The crystal structure of one of the isozymes of OYE has been solved and we have been mutating significant residues to determine the effect on ligand binding and NADPH oxidation by the enzyme. We plan to use several of the features of the GCG program to design primers for additional mutation. The general focus of the lab is the study of riboflavin-containing enzymes. We are currently working on the cloning and characterization of two of these enzymes, D-Amino Acid Oxidase and 2-Methyl-3-Hydroxypyridine-5-Carboxylic Acid Oxygenase. In these studies we will continue to use the features of the GCG package.