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

这项由普林斯顿大学的Jannette Carey和南波希米亚大学的Ivana Smatanova （Ceske Budejovice）合作的美国-捷克研究项目的目标是扩展我们对一类重要蛋白质的结构和分子功能之间关系的认识，并将这些特征与它们的生物学作用联系起来。该项目特别涉及到能弱结合黄素单核苷酸（FMN）的蛋白WrbA的结构研究。这种蛋白存在于多种细菌和真核生物中，属于一个保守的新型多聚体黄毒毒素样蛋白家族。研究人员已经获得了apo-WrbA的衍射质量晶体。对这种蛋白的兴趣源于计算机建模和生化实验的结果，这些结果与同源黄酮毒素相比，建立了有趣的差异，并表明与氧化应激反应有关。这个由美国和捷克的高级和初级生物化学家组成的团队计划：1)对野生型蛋白质进行结构表征；2)对亚结构域洗牌产物进行表征。研究结果有望为辅助因子结合、二聚化和功能域的结构安排提供三维的见解。成功将增强我们对黄素和DNA结合域如何在单个多肽中结合的认识，对指导未来通过定点诱变进行蛋白质工程具有重要意义。该项目是生物学和物理科学的结合，通过使美国和中欧的专家能够在共同感兴趣和能力强的领域结合互补的人才和共享研究资源，实现了推进科学知识的计划目标。更广泛的影响包括通过在合作机构的工作和直接参与高级结构生物学，包括建模技术和x射线衍射分析，将美国和捷克学生引入国际研究界。该项目由美国国家科学基金会分子和细胞生物科学部的生物分子系统集群和国际科学与工程办公室共同资助。

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.