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的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 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组，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，与帕金森病和某些类型的癌症有关。 DJ-1超家族的许多其他成员已被部分表征，并在各种生物对环境胁迫的反应中发挥重要作用。 我们有兴趣确定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实现非常不同的生物学作用。 我们正在研究荧光假单胞菌异腈水合酶和几种细菌一般应激反应蛋白的YhbO家庭。异腈水合酶催化各种异腈转化为酰胺，并使用DJ-1超家族中保守的氨基酸来实现这种独特的化学反应。 YhbO是一个大的和无所不在的一类原核蛋白，保护细菌免受一般的非生物胁迫。 我们已经收集了异腈水合酶的晶体和几个YhbO同系物从各种细菌在UNL的旋转阳极源上的数据集。基于它们的衍射，我们相信这些样品中的每一个都将在APS下达到原子分辨率（dmin < 1.2）。我们已经生长了每一种的多个晶体，并将底物浸入异腈水合酶的催化失活点突变体的晶体中。 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收集的改进数据将有助于模型的改进、分析和出版。

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.