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

加州大学旧金山分校和加州大学圣巴巴拉分校的罗德尼·K·盖伊、布拉德福德·吉布森和欧文·昆茨得到了特别项目办公室化学部的联合支持，以表彰他们在质谱学确定蛋白质结构方面的跨学科合作。他们将设计和合成交联剂，当它们在一定距离内时，这些交联剂将选择性地结合到氨基酸侧链对上。通过单独或组合使用各种交联剂，它们将能够以化学方式连接相距5-20埃的残基。同位素标记的交联剂或包含亲和纯化标签的交联剂也将被研究。修饰后的蛋白质经高效液相色谱消化、纯化后，再进行质谱分析。将开发计算技术来分析质谱学数据并得出蛋白质的三级结构。最初要研究的蛋白质包括成纤维细胞生长因子-2和HIV整合酶。这种方法还将用于研究蛋白质与蛋白质或结构域之间的相互作用。确定蛋白质或蛋白质复合体的三级或三维结构是一个关键问题。这一质谱学工具将为现有的核磁共振和单晶X射线衍射技术提供补充。其目标是开发一种高度自动化、高通量的系统，可以使用少量的蛋白质来推断三级结构。化学合作研究(CRC)奖授予致力于广泛化学兴趣问题的跨学科科学家团队。这些奖项的重点是研究和培训的新合作模式。

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.