Kinome-Wide Spectroscopic Study of Drug Binding Site Electrostatics

项目摘要/摘要 蛋白激酶在调节生长和增殖的信号通路中起着核心作用。 而异常的激酶活性会导致许多癌症的发生。近期 成功地使用靶向蛋白激酶抑制剂治疗特定癌症，特别是肺癌和 慢性髓系白血病，强调了这些蛋白质在肿瘤发生中的重要性，以及 强调需要更多的激酶抑制剂来治疗其他癌症。新技术的发展 激酶抑制剂是具有挑战性的，因为激酶ATP结合的高度序列保守性 作为这些小分子靶标的主要位点，很难获得具有 对特定的激酶有选择性。目前的研究旨在通过一个完全 利用物理化学的进步的新的实验方法。在Aim 1中，一个新的 被称为振动斯塔克光谱的光谱技术将被用来构建 ATP结合位点的静电学及其在该蛋白质的~500个成员中的变化 一家人。这些测量将使用拥有振动探针的激酶抑制剂进行 电场，其中探针报告它们在结合在ATP中时所经历的静电- 结合部位。因为这些静电图与ATP中的物理环境有关- 从抑制剂的角度出现结合位点，他们将直接洞察如何 抑制剂的化学结构的改变会影响与激酶的相互作用。 在这些测量中发现的激酶之间的差异可以被用来设计更多 选择性药物。在目标2中，这种可能性将通过执行大规模的结合分析来量化。 这些激酶抑制剂的选择性将被揭示，并直接与 静电测量，以揭示静电变化如何决定选择性。而选择性 剖面图在制药行业中很常见，与静电图的比较 在目标1中确定将允许第一次确定抑制剂选择性的物理基础 时间，为开发具有新的选择性曲线的缓蚀剂指明了方向。在目标3中， 三磷酸腺苷结合位点的表征将通过研究这种环境是如何完成的 受蛋白质基团和结合水分子动态重排的影响。这种蛋白质 激酶现在构成了一组主要的药理靶点，这项工作将结合在一起 首次对这些蛋白质的物理性质进行了全面的实验研究 决定了它们与药物分子的相互作用。

Project Summary / Abstract Protein kinases play central roles in the signaling pathways that regulate the growth and proliferation of cells, and aberrant kinase activity contributes to the development of many cancers. Recent success in treating particular cancers with targeted protein kinases inhibitors, notably lung cancer and chronic myeloid leukemia, underscores the importance of these proteins in oncogenesis, and highlights the need for additional kinase inhibitors to treat other cancers. The development of new kinase inhibitors is challenging because the high sequence conservation of the kinase ATP-binding site, the major site targeted by these small molecules, makes it difficult to obtain compounds that are selective for particular kinases. The current study aims to address this problem through an entirely new experimental approach that utilizes advances in physical chemistry. In Aim 1, a new spectroscopic technique called vibrational Stark spectroscopy will be used to construct a map of the electrostatics of the ATP-binding site and how it varies across the ~500 members of this protein family. These measurements will be made using kinase inhibitors that possess vibrational probes of electric field, in which the probes report on the electrostatics they experience when bound in the ATP- binding site. Because these electrostatic maps relate to how the physical environment in the ATP- binding site appears from the perspective of the inhibitors, they will yield direct insight into how changes to the chemical structure of the inhibitors would affect the interaction with kinases. Differences uncovered between kinases in these measurements could be exploited to design more selective drugs. In Aim 2, this possibility will be quantified by performing large-scale binding assays in which the selectivity of panels of kinase inhibitors will be revealed and directly compared to the electrostatics measurements to reveal how electrostatic variation dictates selectivity. While selectivity profiling is commonplace in the pharmaceutical industry, the comparison with the electrostatic maps determined in Aim 1 will allow the physical basis of inhibitor selectivity to be determined for the first time, guiding the way to the development of inhibitors with new selectivity profiles. In Aim 3 the characterization of the ATP-binding site will be completed by studying how this environment is affected by the dynamic rearrangements of protein groups and bound water molecules. The protein kinases now constitute a major group of pharmacological targets, and taken together this work will constitute the first comprehensive experimental study of how the physical properties of these proteins dictate their interaction with drug molecules.