Collaborative Research in Chemistry (CRC): Moderate Resolution Protein Structures by Chemical Crosslinking and Mass Spectrometry
Collaborative Research in Chemistry (CRC): Moderate Resolution Protein Structures by Chemical Crosslinking and Mass Spectrometry
批准号:
0118481
负责人:
Irwin Kuntz
金额:
$195.0万
依托单位国家:
美国
项目类别:
Continuing Grant
财政年份:
2001
资助国家:
美国
项目状态:
已结题
起止时间:
2001-09-01 至 2007-08-31
中文摘要
加州大学旧金山分校和加州大学圣塔芭芭拉分校的 Rodney K. Guy、Bradford Gibson 和 Irwin Kuntz 在通过质谱测定蛋白质结构方面开展跨学科合作,得到了化学部特别项目办公室的共同支持。他们将设计和合成交联剂,当氨基酸侧链对在一定距离内时,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多种交联剂,它们将能够以化学方式连接相距 5-20 埃的残基。还将研究同位素标记的交联剂或掺入亲和纯化标签的交联剂。交联后,修饰的蛋白质将被消化,通过 HPLC 纯化,然后通过质谱分析。将开发计算技术来分析质谱数据并推导蛋白质的三级结构。最初要研究的蛋白质包括 FGF-2 和 HIV 整合酶。该方法还将用于研究蛋白质-蛋白质或域-域相互作用。确定蛋白质或蛋白质复合物的三级或三维结构是一个关键问题。该质谱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。目标是开发一种高度自动化、高通量的系统,可以使用少量蛋白质推断三级结构。化学合作研究 (CRC) 奖项颁发给致力于解决广泛化学问题的跨学科科学家团队。这些奖项的重点是新的研究和培训合作模式。
英文摘要
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.
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Isolation and Characterization of BPTI Folding Mutants
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批准号:8606901
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项目类别:Continuing grant
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资助金额:$0.0万
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财政年份:1986
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负责人:Irwin Kuntz
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项目类别:Standard Grant
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资助金额:$0.0万
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财政年份:1978
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负责人:Irwin Kuntz
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依托单位:
国内基金
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