STRUCTURAL STUDIES OF THE EVOLUTION OF NEW FUNCTION IN THE DJ-1 SUPERFAMILY

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 DJ-1超家族新功能进化的结构研究 DJ-1超家族是一个庞大而多样的蛋白质集合，在大多数生命王国中都有代表。人类DJ-1是一种保护细胞免受氧化应激的蛋白质，与帕金森氏症、S病和某些类型的癌症有关。DJ-1超家族的许多其他成员已经被部分描述，并在各种生物对环境胁迫的反应中发挥重要作用。我们感兴趣的是确定DJ-1超家族中相关但功能不同的蛋白质如何使用相似的结构特征来履行非常不同的生物角色。 我们正在研究YhbO家族中的假瘤荧光异腈水合酶和几种细菌一般应激反应蛋白。异腈水合酶催化各种异腈转化为酰胺，并利用DJ-1超家族中保守的氨基酸来完成这一独特的化学作用。YhbO是一大类无处不在的原核蛋白，可以保护细菌免受一般非生物胁迫的影响。我们收集了UNL旋转阳极源上各种细菌的异腈水合酶晶体和几个YhbO同系物的数据集。根据它们的衍射，我们相信每个样品都将在APS上衍射到原子分辨率(dmin&lt；1.2？)。我们已经生长了每个异腈水合酶的多个晶体，并将底物浸泡到异腈水合酶的催化失活点突变的晶体中。这些结构中的每一个都已经通过分子置换成功地解决了，我们将在APS收集的改进的数据将有助于模型的完善、分析和发表。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Structural Studies of the Evolution of New Function in the DJ-1 Superfamily The DJ-1 superfamily is a large and diverse set of proteins that has representatives in most kingdoms of life. Human DJ-1 is protein that protects cells against oxidative stress, and in implicated in both Parkinson¿s disease and certain types of cancer. Many other members of the DJ-1 superfamily have been partially characterized and play important roles in the response of various organisms to environmental stress. We are interested in determining how related but functionally distinct proteins in the DJ-1 superfamily use the similar structural features to fulfill very different biological roles. We are studying Pseudomomas fluorescens isonitrile hydratase and several bacterial general stress response proteins in the YhbO family. Isonitrile hydratase catalyzes the conversion of various isonitriles to amides and employs amino acids that are well-conserved in the DJ-1 superfamily to accomplish this unique chemistry. YhbO is a large and omnipresent class of prokaryotic proteins that protect bacteria from general abiotic stresses. We have collected datasets on crystals of isonitrile hydratase and several YhbO homologues from various bacteria on a rotating anode source at UNL. Based on their diffraction, we believe that each of these samples will diffract to atomic resolution (dmin < 1.2 ¿) at the APS. We have grown multiple crystals of each and have soaked substrate into crystals of a catalytically inactive point mutant of isonitrile hydratase. Each of these structures has been successfully solved by molecular replacement and the improved data that we will collect at the APS will aid in model refinement, analysis, and publication.