MECHANISM OF PROMISCUOUS INHIBITION BY SMALL MOLECULE AGGREGATION

小分子聚集的混杂抑制机制

• 批准号: 8169763
• 负责人: Brian K Shoichet
• 金额: $ 0.18万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-12 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169763
• 关键词: Address; Amides; Behavior; Binding; Biological; Communities; Computer Retrieval of Information on Scientific Projects Database; Coupled; Deuterium; Enzymes; Funding; Grant; Hydrogen; Institution; Lead; Mass Spectrum Analysis; Membrane Proteins; Motion; Proteins; Protons; Research; Research Personnel; Resources; Solvents; Source; Surface; Techniques; United States National Institutes of Health; inhibitor/antagonist; insight; novel; protein aggregate; small molecule

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The scientific community has long been troubled by the presence of nonspecific small molecules that inhibit proteins other than their hypothesized targets. The behavior of a large and diverse subset of these inhibitors can be explained by a common mechanism: aggregation. At micromolar concentrations some nonspecific molecules form large aggregates that inhibit a wide range of unrelated soluble proteins. From previous studies we know that inhibition results from the direct interaction of protein and aggregates. This interaction is reversible and does not result in denaturation, but the extent of our understanding stops there. How does binding to the aggregate lead to inhibition? Does the aggregate bind to a particular protein surface or is it nonspecific? Does binding restrain or induce dynamic motion in the protein? I will use deuterium exchange mass spectrometry to address these questions.Since we know that aggregates induce changes subtler than denaturation, it is necessary that we use a more sensitive technique. Mass spectrometry coupled to deuterium exchange of amide hydrogens provides evidence of how exposed certain regions are to solvent. This allows us to examine the accessibility of surface protons and the overall dynamics of the enzyme through the exchange of core (or buried) protons. We predict that aggregates inhibit by altering the normal dynamic motions of the enzyme. Additionally, we will be able to determine whether there is a particular surface to which the aggregate binds. There are few techniques that allow this level of sensitivity and among them mass spectrometry is the most practical. These studies will provide insight into the mechanism of promiscuous inhibition by aggregation, thus elucidating a novel interaction between small molecules and their biological targets.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 长期以来，科学界一直对非特异性小分子的存在感到困扰，这些小分子会抑制其假设目标以外的蛋白质。 这些抑制剂的大量且多样化的子集的行为可以通过一个共同的机制来解释：聚集。 在微摩尔浓度下，一些非特异性分子形成大的聚集体，抑制多种不相关的可溶性蛋白质。 从之前的研究中我们知道抑制是蛋白质和聚集体直接相互作用的结果。 这种相互作用是可逆的，不会导致变性，但我们的理解范围仅限于此。 与聚集体的结合如何导致抑制？聚集体是否与特定的蛋白质表面结合还是非特异性的？ 结合是否抑制或诱导蛋白质的动态运动？我将使用氘交换质谱来解决这些问题。由于我们知道聚集体引起的变化比变性更微妙，因此我们有必要使用更灵敏的技术。 与酰胺氢的氘交换相结合的质谱分析提供了某些区域如何暴露于溶剂的证据。 这使我们能够通过核心（或埋藏）质子的交换来检查表面质子的可及性和酶的整体动力学。 我们预测聚集体通过改变酶的正常动态运动来抑制。 此外，我们将能够确定是否存在聚集体结合的特定表面。 很少有技术可以达到这种水平的灵敏度，其中质谱法是最实用的。 这些研究将深入了解聚集的混杂抑制机制，从而阐明小分子与其生物靶标之间的新型相互作用。