Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX 77204, United States.
Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX 77204, United States.
Biochim Biophys Acta Gen Subj. 2017 Nov;1861(11 Pt A):2640-2651. doi: 10.1016/j.bbagen.2017.07.006. Epub 2017 Jul 13.
Resveratrol (1) is a naturally occurring polyphenol that has been implicated in neuroprotection. One of resveratrol's several biological targets is Ca-sensitive protein kinase C alpha (PKCα). Resveratrol inhibits PKCα by binding to its activator-binding C1 domain. Munc13-1 is a C1 domain-containing Ca-sensitive SNARE complex protein essential for vesicle priming and neurotransmitter release.
To test if resveratrol could also bind and inhibit Munc13-1, we studied the interaction of resveratrol and its derivatives, (E)-1,3-dimethoxy-5-(4-methoxystyryl)benzene, (E)-5,5'-(ethene-1,2-diyl)bis(benzene-1,2,3-triol), (E)-1,2-bis(3,4,5-trimethoxyphenyl)ethane, and (E)-5-(4-(hexadecyloxy)-3,5-dihydroxystyryl)benzene-1,2,3-triol with Munc13-1 by studying its membrane translocation from cytosol to plasma membrane in HT22 cells and primary hippocampal neurons.
Resveratrol, but not the derivatives inhibited phorbol ester-induced Munc13-1 translocation from cytosol to membrane in HT22 cells and primary hippocampal neurons, as evidenced by immunoblot analysis and confocal microscopy. Resveratrol did not show any effect on Munc13-1, a mutant which is not sensitive to phorbol ester. Binding studies with Munc13-1 C1 indicated that resveratrol competes with phorbol ester for the binding site. Molecular docking and dynamics studies suggested that hydroxyl groups of resveratrol interact with phorbol-ester binding residues in the binding pocket.
This study characterizes Munc13-1 as a target of resveratrol and highlights the importance of dietary polyphenol in the management of neurodegenerative diseases.
白藜芦醇（1）是一种天然存在的多酚，已被牵涉到神经保护中。白藜芦醇的几个生物靶点之一是钙敏感蛋白激酶 Cα（PKCα）。白藜芦醇通过结合其激活剂结合 C1 结构域来抑制 PKCα。Munc13-1 是一种含有 C1 结构域的钙敏感 SNARE 复合蛋白，对于囊泡引发和神经递质释放至关重要。
为了测试白藜芦醇是否也可以结合并抑制 Munc13-1，我们研究了白藜芦醇及其衍生物（E）-1,3-二甲氧基-5-（4-甲氧基苯乙烯基）苯、（E）-5,5' -（乙烯-1,2-二基）双（苯-1,2,3-三醇）、（E）-1,2-双（3,4,5-三甲氧基苯基）乙烷和（E）-5-（4-（十六烷氧基）-3,5-二羟基苯乙烯基）苯-1,2,3-三醇与 Munc13-1 的相互作用，通过研究其在 HT22 细胞和原代海马神经元中从细胞质向质膜的膜易位。
白藜芦醇，但不是衍生物，抑制佛波醇酯诱导的 Munc13-1 从 HT22 细胞和原代海马神经元的细胞质向膜的易位，如免疫印迹分析和共聚焦显微镜所示。白藜芦醇对 Munc13-1 没有任何作用，Munc13-1 对佛波醇酯不敏感。用 Munc13-1 C1 进行的结合研究表明，白藜芦醇与佛波醇酯竞争结合位点。分子对接和动力学研究表明，白藜芦醇的羟基与结合口袋中佛波醇结合残基相互作用。
本研究将 Munc13-1 鉴定为白藜芦醇的靶点，并强调了膳食多酚在神经退行性疾病管理中的重要性。
