Kinome-Wide Spectroscopic Study of Drug Binding Site Electrostatics

项目总结/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 激酶活性异常导致许多癌症的发生。最近 用靶向蛋白激酶抑制剂治疗特定癌症，特别是肺癌的成功， 慢性髓性白血病，强调了这些蛋白质在肿瘤发生中的重要性， 强调了需要额外的激酶抑制剂来治疗其他癌症。发展新 激酶抑制剂具有挑战性，因为激酶ATP结合的高序列保守性 位点，这些小分子靶向的主要位点，使得难以获得 对特定的激酶有选择性。目前的研究旨在通过一个全面的 利用物理化学进步的新实验方法。在目标1中， 一种称为振动斯塔克光谱的光谱技术将被用来构建一个地图， 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 家人这些测量将使用具有以下振动探针的激酶抑制剂进行： 电场，其中探针报告它们在ATP中结合时所经历的静电- 结合位点因为这些静电地图与ATP中的物理环境有关- 从抑制剂的角度来看，结合位点的出现，他们将直接了解如何 抑制剂化学结构的改变将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用来设计更多的 选择性药物在目标2中，这种可能性将通过进行大规模的结合试验来量化， 这将揭示激酶抑制剂组的选择性，并直接与 静电测量，以揭示静电变化如何决定选择性。虽然选择性 剖析是司空见惯的制药工业，比较与静电地图 目标1中确定的抑制剂选择性的物理基础将允许确定第一个 时间，指导方式的发展与新的选择性的抑制剂。在Aim 3中， ATP结合位点的表征将通过研究这种环境是如何 受到蛋白质组和结合水分子的动态重排的影响。蛋白质 激酶现在构成了一组主要的药理学靶点，综合起来，这项工作将 构成了第一个全面的实验研究，这些蛋白质的物理性质， 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.