Optimization of modulators of Gbg-SNARE interaction

Gbg-SNARE 相互作用调制器的优化

• 批准号: 9085379
• 负责人: HEIDI E HAMM
• 金额: $ 39.14万
• 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 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9085379
• 关键词: Acetylcholine; Address; Affect; Agonist; Binding; Bioavailable; Biological Assay; Biological Availability; Brain; Cells; Chemicals; Complex; Coupled; Cytoplasmic Granules; Development; Disease; Distal; Dopamine; Electrophysiology (science); Enhancers; Event; Exocytosis; Functional disorder; Future; G-Protein-Coupled Receptors; G-protein Beta gamma; 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; 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; neuropsychiatric disorder; neuroregulation; neurotransmission; neurotransmitter release; novel; novel therapeutics; 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（由适用提供）：通过受调节的胞吐作用释放化学发射器是许多形式的细胞间通信，包括骑马的释放和突触传播。胞吐作用受到复杂的调节，涉及蛋白质 - 蛋白质相互作用和膜重塑事件的网络。 G蛋白偶联受体（GPCR）在策划这一复杂调节中起着核心作用，而GI/O-偶联的GPCR众所周知，通过释放G蛋白βγ亚基，可以抑制神经分泌细胞从神经分泌细胞中释放发射机。这种深刻的抑制有可能有助于突触前整合和合成可塑性。这种抑制作用的最佳研究机制是调节Ca2+通道的电压灵敏度。然而，Gβγ还可以通过与固体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复合物结合来直接抑制Ca2+进入点的神经递质释放。我们已经表明，SNAP25的C端对于抑制发射机释放的能力至关重要。我们设计了一个测定法 Gβγ-SNAP25使用Alphascreen的相互作用，并筛选了一个基于已知的化学型设计的小型化合物库，这些化合物是根据已知的化学型设计的，这些化学型调节了抑制剂和增强剂的蛋白质蛋白质相互作用（PPI）。在这笔赠款中，我们将 在AIM 2中，我们将确定与其他Gβγ相互作用蛋白相比Gβγ-SNAP25相互作用的化合物的选择性。在AIM 3中，我们将确定优化分子对培养中海马神经元释放神经递质的影响，并确定它们是否与突触前GI/O偶联GPCR的激动剂和拮抗剂合成。这些化合物应使我们能够评估Gβγ-SNARE相互作用对GPCR调节胞吐作用的重要性。它们可能与突触前GPCR的激动剂或拮抗剂并行工作，从而与突触前功能协同作用，有选择地影响神经递质的突触前但没有突触后作用。我们提出的研究解决了神经科学的基本问题，神经递质控制胞吐作用的分子机制。这些研究可能定义了开发新疗法的靶标，这些靶标可能会揭示与分泌和神经调节有关的疾病的病理基础。

项目成果

Specificities of Gβγ subunits for the SNARE complex before and after stimulation of α2a-adrenergic receptors.

刺激 α2a 肾上腺素能受体之前和之后 SNARE 复合体的 Gβγ 亚基的特异性。

• DOI: 10.1126/scisignal.abc4970
• 发表时间: 2021
• 期刊: Science signaling
• 影响因子: 7.3
• 作者: Yim,YunYoung;McDonald,WHayes;Betke,KatherineM;Kaya,Ali;Hyde,Karren;Erreger,Kevin;Gilsbach,Ralf;Hein,Lutz;Hamm,HeidiE
• 通讯作者: Hamm,HeidiE