Collaborative Research in Chemistry (CRC): Moderate Resolution Protein Structures by Chemical Crosslinking and Mass Spectrometry

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Rodney K. Guy, Bradford Gibson and Irwin Kuntz of the University of California San Francisco and the University of California Santa Barbara are jointly supported by the Division of Chemistry, Special Projects Office, for their interdisciplinary collaboration on protein structure determinations by mass spectrometry. They will design and synthesize cross-linking agents that will selectively bind to pairs of amino acid sidechains when they are within a certain distance. By using a variety of crosslinking agents singly or in combination, they will be able to chemically connect residues that are 5-20 Angstroms apart. Isotopically labeled crosslinkers or crosslinkers that incorporate an affinity purification tag will also be investigated. After crosslinking, the modified proteins will be digested, purified by HPLC, then analyzed by mass spectrometry. Computational techniques will be developed to analyze the mass spectrometric data and derive the tertiary structure of the protein. Initial proteins to be studied include FGF-2 and HIV-integrase. This method will also be used to study protein-protein or domain-domain interactions.Determining the tertiary or three dimensional structure of proteins or protein complexes is a key problem. This mass spectrometric tool will provide a complement to existing techniques of nuclear magnetic resonance and single crystal x-ray diffraction. The goal is to develop a highly automated, high-throughput system that can infer tertiary structure using small amounts of protein.Collaborative Research in Chemistry (CRC) awards are given to interdisciplinary teams of scientists working on problems of broad chemical interest. The emphasis in these awards is on new collaborative modes of research and training.