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

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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.