The Center for Eukaryotic Structural Genomics
The Center for Eukaryotic Structural Genomics is a collaborative effort to elucidate the three-dimensional structures of many of the proteins encoded by the Arabidopsis thaliana genome. Starting with genomic DNA or a cDNA library, selected targets are cloned into a vector for bacterial or wheat germ cell-free protein expression, from which recombinant protein is subsequently purified. The three dimensional structure of the proteins is then determined via either X-ray crystallography or nuclear magnetic resonance (NMR). All data and solved structures are deposited in public databases including the Protein Data Bank (PDB) and BioMagResBank (BMRB).
The CESG is primarily located in the Department of Biochemistry at the University of Wisconsin-Madison with a significant effort in NMR structural analysis in our laboratory at the Medical College of Wisconsin here in Milwaukee. Other partners include Tokyo Metropolitan University (Japan) and Hebrew University (Israel).
Our role in the CESG is to develop and apply streamlined methods for the acquisition and analysis of NMR data for the determination of three-dimensional protein structures. With a cryoprobe-equipped 600 MHz NMR spectrometer we collect complete structural datasets in a fraction of the time required with conventional instrumentation. When coupled with new software tools designed to automate most of the routine tasks in the data analysis pathway, we hope to contribute to a rapid maturation in the standard methodology for structural protein NMR.
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NMR structure of the hypothetical protein At3g17210 from the model plant system Arabidopsis thaliana
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The first protein structure determined by the CESG was recently completed in our laboratory by Dr. Betsy Lytle and Dr. Francis Peterson. At3g17210 is a hypothetical protein cloned from the Arabidopsis genome based on a combination of selection criteria designed to bias our efforts toward novel folds and proteins of unknown structure and function. The homodimeric structure observed for At3g17210 represents a novel fold that was recently observed for the first time in a bacterial monooxygenase protein, ActVA-Orf6, from Streptomyces coelicolor. These results demonstrate the need for structural genomics initiatives in populating the protein fold universe densely enough to enable reliable modeling of the new sequences emanating from genome sequencing projects.
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