U.S.-Czech Biomolecular Research on Structural Studies of a Novel Flavodoxin-like Protein

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The goal of this U.S.-Czech research project between Jannette Carey of Princeton University and Ivana Smatanova of the University of South Bohemia, Ceske Budejovice, is to extend our knowledge of the relationships between structure and molecular functions in an important class of proteins and to relate these features to their biological roles. The project specifically involves the structural investigation of the protein WrbA, which can weakly bind flavin mononucleotide (FMN). This protein is found in diverse bacteria and eukaryotes and belongs to a conserved family of novel multimeric flavodoxin-like proteins. The researchers have obtained diffraction-quality crystals of the apo-WrbA. Interest in this protein stems from available results from computer modeling and biochemical experiments that establish interesting differences when compared with homologous flavodoxins and suggest involvement in oxidative-stress responses. The team of senior and junior U.S. and Czech biochemists plans to: 1) structurally characterize the wild type protein and 2) characterize subdomain-shuffling products. Results are expected to provide three-dimensional insights into the architectural arrangement of the cofactor binding, dimerization, and functional domains. Success would enhance our knowledge of how flavin and DNA binding domains are combined in a single polypeptide, with implications for guiding future protein engineering through site-directed mutagenesis. This project at the interface of biology and physical sciences fulfills the program objective of advancing scientific knowledge by enabling experts in the United States and Central Europe to combine complementary talents and share research resources in areas of strong mutual interest and competence. Broader implications include the introduction of U.S. and Czech students to the international research community through work at partner institutions and direct involvement in advanced structural biology, including modeling techniques and x-ray diffraction analysis. The project is funded jointly by the Biomolecular Systems Cluster in the Division of Molecular and Cellular Biosciences and the Office of International Science and Engineering at NSF.