The Nuclear Magnetic Resonance (NMR) Center is used to study the three-dimensional structure of biological molecules, including proteins, nucleic acids, carbohydrates and lipids. The research focuses primarily on protein domains involved in endocytosis,
signal transduction, viral replication, and neurotransmission.
The first three-dimensional structure of an Eps15 homology domain bound to an endocytic peptide signal has been solved in the NMR Center. It reveals how the Eps15 protein contributes to the regulation of cell growth. The structure of a domain of the EEA1 protein bound to a phospholipid has been determined, revealing how membrane lipids are specifically recognized. Proteins that package viral DNA and bind alcohol and neurotransmitters are also being characterized. Understanding the structural mechanisms of these proteins will inform the design of new therapeutic agents for cancer, Parkinson's disease, and alcohol toxicity and addiction.
For the past three decades, the Department of Pharmacology at the University of Colorado School of Medicine has maintained a presence in the advancement of mass spectrometric techniques to solve difficult problems in the pharmacological sciences.
In the past, this has taken form in the elucidation of chemical structures of biologically active lipids derived from arachidonic acid and delineation of the biochemical mechanisms involved in the metabolism of these important eicosanoids. Mass spectrometry has also been a central tool to expand our understanding of the complexities of eicosanoid biosynthesis within cells in tissues.However, with the post-genomic era, mass spectrometry has now emerged as a central technique that permits precise identification of proteins expressed within cells. Investigators in the Department of Pharmacology and colleagues in the Department of Pharmaceutical Sciences (School of Pharmacy) have installed the new generation of mass spectrometric instruments, including:
One example of current capability is the study of the covalent binding of leukotriene A4 with leukotriene A4 hydrolase and the isolation of a specific peptide containing the covalently linked leukotriene A4 (1). Detailed mass spectrometric investigation
permitted the sequencing of the peptide and the assignment of the covalent attachment site as tyrosine-383 close to the active site of leukotriene A4 hydrolase. Such detailed biochemical information provides some insight into the enzymatic mechanism
of LTA, hydrolase as well as a broader understanding of potential agents which may interfere with the production of leukotriene B, and thus, pharmacologically control this enzyme.
The Biomolecular X-ray Crystallography Center provides the facilities to the university community for production of crystals of macromolecules and determination of high resolution 3-dimensional structures using X-ray crystallography. For example, whether genes
are regulated directly by proteins in transcription complexes or indirectly through signals generated by bacterial quorum sensing systems, structural analysis is an essential and illustrative approach to understanding function and developing therapeutic
agents. Users of the X-ray Crystallography Center are also interested in the way that molecular recognition is involved in a variety of processes.
Current projects include the study of chromosomal protein-DNA complexes, structural studies of tumor viruses and viral protein interactions, DNA modification enzymes, and proteins involved in bacterial pathogenesis. Learning more about the structure and function of the molecules active in these systems will advance our understanding of fundamental molecular processes in the cell and aid in fighting human disease.
PhD, 2009, Columbia University
Molecular Mechanisms of ion channel function. Examining structural and regulatory mechanisms of the Acid-sensing ion channels using electrophysiology, fluorescence, spectroscopy, and structural biology.
Churchill, Mair E. A.
PhD, 1987, Johns Hopkins Univ.
Structure and mechanism in gene regulation; biophysical and structural studies of protein-nucleic acid and protein-protein complexes in chromatin and bacterial pathogenesis.
Jones, David N.M.
PhD, 1989, Univ. of Cambridge
Molecular mechanism of alcohols and anesthetic actions; structure and function of biomolecules; NMR spectroscopy, x-ray crystallography, biophysics and molecular biology.
Kieft, Jeffrey S.
PhD, 1997, Univ. of California, Berkeley
The way by which viral RNAs, with their diverse and dynamic structures, can hijack the machinery of an infected cell and using this information to understand basic biological processes.
Kutateladze, Tatiana G.
PhD, 1988, Moscow State Univ.
Epigenetics, phosphoinositide signaling, structural biology, NMR and crystal structures of proteins implicated in cancer, structure based drug design.