Optimization of modulators of Gbg-SNARE interaction

Gbg-SNARE 相互作用调制器的优化

• 批准号: 8856366
• 负责人: HEIDI E HAMM
• 金额: $ 39.41万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8856366
• 关键词: Acetylcholine; Address; Affect; Agonist; Binding; Bioavailable; Biological Assay; Biological Availability; Brain; Cells; Chemicals; Complex; Coupled; Cytoplasmic Granules; Development; Disease; Distal; Dopamine; Enhancers; Event; Exocytosis; Functional disorder; Future; G-Protein-Coupled Receptors; GTP-Binding Proteins; Grant; Health; Hippocampus (Brain); Hormones; Image; In Vitro; Internet; Investigation; Learning; Libraries; Light; Mediating; Membrane; Memory; Molecular; Neuraxis; Neurons; Neurosciences; Neurotransmitter Receptor; Neurotransmitters; Norepinephrine; Pathology; Phosphatidylinositols; Phosphotransferases; Physiological; Play; Proteins; Regulation; Regulation of Exocytosis; Relative (related person); Reporting; Role; SNAP receptor; Second Messenger Systems; Serotonin; Site; Slice; Specificity; Structure-Activity Relationship; Synaptic Transmission; Synaptic Vesicles; Synaptic plasticity; System; Testing; Therapeutic; Work; base; chemical release; design; high throughput screening; in vivo; information processing; inhibitor/antagonist; intercellular communication; millisecond; neuropsychiatry; neuroregulation; neurotransmission; neurotransmitter release; novel; postsynaptic; presynaptic; presynaptic neurons; protein protein interaction; receptor; receptor binding; receptor coupling; second messenger; sensor; small molecule; synaptotagmin; tool; voltage

DESCRIPTION (provided by applicant): Release of chemical transmitters by regulated exocytosis underlies many forms of intercellular communication, including hormone release and synaptic transmission. Exocytosis is subject to complex modulation and involves a web of protein-protein interactions and membrane remodeling events. G protein-coupled receptors (GPCRs) play a central role in orchestrating this complex regulation, and Gi/o- coupled GPCRs are well known to inhibit transmitter release from neurosecretory cells by release of G protein βγ subunits. This profound inhibition has the potential to contribute to presynaptic integration and synaptic plasticity. The best-studied mechanism for this inhibition is modulation of the voltage sensitivity of Ca2+ channels. However, Gβγ can also directly inhibit neurotransmitter release at a point distal to Ca2+ entry by binding to soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins as well as the assembled SNARE complex. We have shown that the C-terminus of SNAP25 is critical for the ability to inhibit transmitter release. We have devised an assay of Gβγ-SNAP25 interaction using the AlphaScreen, and have screened a small library of compounds that were designed based on known chemotypes which modulate protein-protein interaction (PPI) for both inhibitors and enhancers of this. In this grant, we will optimize both inhibitors and enhancers of Gβγ-SNAP25 interaction In Aim 1, we will carry out medicinal chemical optimization of both classes of PPI compounds to increase their potency, selectivity, and bioavailability. In Aim 2, we will determine the selectivity of the compounds for Gβγ-SNAP25 interaction compared to other Gβγ-interacting proteins. In Aim 3, we will determine the effects of the optimized molecules on neurotransmitter release from hippocampal neurons in culture, and determine whether they synergize with the agonists and antagonists of presynaptic Gi/o-coupled GPCRs. These compounds should allow us to evaluate the importance of Gβγ-SNARE interaction for GPCR modulation of exocytosis. They may work in parallel with agonists or antagonists of presynaptic GPCRs, and thus synergize with presynaptic functions, selectively affecting presynaptic but not postsynaptic actions of neurotransmitters. The investigations we propose address a fundamental problem in neuroscience, the molecular mechanisms by which neurotransmitters control exocytosis. These studies may define targets for the development of new therapies that may shed light on the pathological basis of diseases related to secretion and neuromodulation.

描述（由申请人提供）：通过受调节的胞吐作用释放化学递质是多种形式的细胞间通讯的基础，包括激素释放和突触传递，胞吐作用受到复杂的调节，并涉及一系列蛋白质-蛋白质相互作用和膜重塑事件。 -耦合受体 (GPCR) 在协调这种复杂的调节中发挥着核心作用，众所周知，Gi/o-耦合 GPCR 可通过释放G 蛋白 βγ 亚基具有促进突触前整合和突触可塑性的潜力。这种抑制的最佳研究机制是调节 Ca2+ 通道的电压敏感性。然而，Gβγ 也可以在某个点直接抑制神经递质释放。我们已经证明，通过与可溶性 N-乙基马来酰亚胺敏感因子附着蛋白受体 (SNARE) 蛋白以及组装的 SNARE 复合物结合，Ca2+ 进入远端。 SNAP25 的 C 末端对于抑制递质释放的能力至关重要。我们设计了一种检测方法。 使用 AlphaScreen 进行 Gβγ-SNAP25 相互作用，并筛选了一个小型化合物库，这些化合物是根据已知的化学型设计的，这些化学型可调节蛋白质-蛋白质相互作用 (PPI) 的抑制剂和增强剂。 优化 Gβγ-SNAP25 相互作用的抑制剂和增强剂 在目标 1 中，我们将对两类 PPI 化合物进行药物化学优化，以提高其效力、选择性和生物利用度 在目标 2 中，我们将确定化合物的选择性。 Gβγ-SNAP25 相互作用与其他 Gβγ 相互作用蛋白的比较 在目标 3 中，我们将确定优化分子对海马神经递质释放的影响。培养中的神经元，并确定它们是否与突触前 Gi/o 偶联 GPCR 的激动剂和拮抗剂协同作用。这些化合物应该使我们能够评估 Gβγ-SNARE 相互作用对于 GPCR 胞吐作用调节的重要性。或突触前 GPCR 的拮抗剂，从而与突触前功能协同作用，选择性地影响突触前而不是突触后的作用我们提出的研究解决了神经科学中的一个基本问题，即神经递质控制胞吐作用的分子机制，这些研究可能会确定新疗法的开发目标，从而揭示与分泌和神经调节相关的疾病的病理基础。