Hydroxyl radical mapping of protein interfaces

描述（由申请人提供）：鉴定a 形成药物结合位点的蛋白质，或存在于药物结合位点的蛋白质。 蛋白质-蛋白质复合物中的界面很难从 单个伴侣蛋白的晶体结构。Carta Proteomics是 致力于通过质量监测同位素交换到蛋白质中的前提 光谱法将允许快速识别接触残留物 形成大分子复合物的蛋白质。以前的工作和专利 发展已经建立了酰胺质子溶剂交换作为Carta的一个方面 蛋白质组学本建议是开发一种羟基互补的方法 自由基（-OH）诱导1 I-TJ 2 H交换到氨基的脂族碳中 酸性侧链。该提案利用了基本的基础 最近已经被很好地表征的化学。 本提案的目的是证明这种能力 使用凝血酶上存在的多个结合位点来验证 该方法正确识别BABIM中活性位点的能力 凝血酶复合物，并确定水蛭素的变构结合位点， 抑制性寡肽。本演示将验证 从组合化学筛选中鉴定抑制剂的方法。 拟定商业应用： 基于结构的药物设计由于缺乏低成本、快速的结构测定方法而受到严重阻碍。我们确定小分子/蛋白质靶相互作用结构的技术可以很容易地以非常低的成本适应高通量和非常快的周转。由于这些独特的功能，我们希望该技术能够很容易地整合到小分子药物发现过程中。Carta Proteomics将通过研究合作协议提供专有专业知识。

DESCRIPTION (provided by applicant): Identifying amino acid residues of a protein that form the binding site for a drug, or that are present at the interface in a protein-protein complex are difficult to identify from the crystal structures of the individual partner proteins. Carta Proteomics is committed to the premise that monitoring isotope exchange into proteins by mass spectrometry will permit the rapid identification of contact residues of proteins that form macromolecular complexes. Prior work and proprietary development has established amide-proton solvent exchange as one facet of Carta Proteomics. This proposal is to develop a complementary method of hydroxyl radical (-OH) induced 1I-TJ2H exchange into the aliphatic carbons of the amino acid side chains. This proposal takes advantage of the basic underlying chemistry that has been recently well characterized. The goal of this proposal is to demonstrate the capability of this chemistry to use the multiple binding sites present on thrombin to validate the ability of this methodology to correctly identify the active site in a BABIM thrombin complex and to identify the allosteric binding site for hirudin, an inhibitory oligopeptide. This demonstration will validate the application of the method to inhibitors identified from combinatorial chemistry screens. PROPOSED COMMERCIAL APPLICATION: Structure based drug design is severely hampered by the lack of a low-cost, rapid method of determining structure. Our technology of determining structures of a small molecule/protein target interaction can be easily adapted to high throughput and very fast turn-around at very low cost. Because of these unique features we expect the technology to be readily integrated into the small molecule drug discovery process. Carta Proteomics will participate by providing the proprietary expertise through research partnership agreements.