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.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 长期以来，科学界一直被非特定小分子的存在所困扰，这些小分子可以抑制蛋白质，而不是它们假设的靶标。这些抑制物中大量且不同的子集的行为可以用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，抑制一系列无关的可溶性蛋白质。从以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，但我们的理解仅限于此。与聚集体的结合是如何导致抑制的？聚集体是结合到特定的蛋白质表面还是非特异性的？结合是否抑制或诱导蛋白质的动态运动？我将使用氢交换质谱仪来解决这些问题。因为我们知道聚集体引起的变化比变性更微妙，所以我们有必要使用更灵敏的技术。质谱学与胺氢的重氢交换相结合，提供了某些区域暴露在溶剂中的证据。这使我们能够通过交换核心(或埋藏的)质子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而受到抑制。此外，我们将能够确定是否存在聚集体绑定到的特定表面。很少有技术可以达到这种程度的灵敏度，其中质谱学是最实用的。这些研究将深入了解聚集抑制混杂的机制，从而阐明小分子与其生物靶标之间的一种新的相互作用。