Wells Christopher A, Betke Katherine M, Lindsley Craig W, Hamm Heidi E
Department of Pharmacology Vanderbilt University Medical Center 442 Robinson Research Building 23rd Ave. South @ Pierce Nashville, Tennessee 37232-6600.
ACS Chem Neurosci. 2012 Jan 18;3(1):69-78. doi: 10.1021/cn200102d.
G(i/o)-coupled presynaptic GPCRs are major targets in neuropsychiatric diseases. For example, presynaptic auto- or heteroreceptors include the D(2) dopamine receptor, H(3) histamine receptor, 5HT(1) serotonin receptors, M(4) acetylcholine receptors, GABA(B) receptors, Class II and III metabotropic glutamate receptors, opioid receptors, as well as many other receptors. These GPCRs exert their influence by decreasing exocytosis of synaptic vesicles. One mechanism by which they act is through direct interaction of the Gβγ subunit with members of the SNARE complex downstream of voltage-dependent calcium channels, and specifically with the C-terminus of SNAP25 and the H3 domain of syntaxin1A(1-3). Small molecule inhibitors of the Gβγ-SNARE interaction would allow the study of the relative importance of this mechanism in more detail. We have utilized novel, label-free technology to detect this protein-protein interaction and screen for several small molecule compounds that perturb the interaction, demonstrating the viability of this approach. Interestingly, the screen also produced enhancers of the Gβγ-SNARE interaction.
G(i/o)偶联的突触前G蛋白偶联受体（GPCRs）是神经精神疾病的主要靶点。例如，突触前自身或异源受体包括D(2)多巴胺受体、H(3)组胺受体、5HT(1)5-羟色胺受体、M(4)乙酰胆碱受体、GABA(B)受体、II类和III类代谢型谷氨酸受体、阿片受体以及许多其他受体。这些GPCRs通过减少突触小泡的胞吐作用发挥其影响。它们起作用的一种机制是通过Gβγ亚基与电压依赖性钙通道下游的SNARE复合体成员直接相互作用，特别是与SNAP25的C末端和Syntaxin1A的H3结构域相互作用（1-3）。Gβγ-SNARE相互作用的小分子抑制剂将有助于更详细地研究这种机制的相对重要性。我们利用了新型的无标记技术来检测这种蛋白质-蛋白质相互作用，并筛选了几种干扰这种相互作用的小分子化合物，证明了这种方法的可行性。有趣的是，筛选还产生了Gβγ-SNARE相互作用的增强剂。
