Resveratrol inhibits phorbol ester-induced membrane translocation of presynaptic Munc13-1

• Published in: Biochimica et biophysica acta Vol. 1861; no. 11; pp. 2640 - 2651
• Main Authors: Pany, Satyabrata, Ghosh, Anamitra, You, Youngki, Nguyen, Nga, Das, Joydip
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.11.2017
• Subjects: Activation; Animals; Binding Sites; confocal microscopy; cytosol; Free Radical Scavengers - administration & dosage; Free Radical Scavengers - chemistry; Hippocampus - drug effects; Hippocampus - metabolism; Humans; Inhibition; Membrane translocation; Mice; Molecular Docking Simulation; molecular models; Munc13-1; mutants; Nerve Tissue Proteins - antagonists & inhibitors; Nerve Tissue Proteins - chemistry; Nerve Tissue Proteins - metabolism; neurodegenerative diseases; neurons; Neurons - drug effects; Neurons - metabolism; Neuroprotection; neuroprotective effect; neurotransmitters; Phorbol Esters - administration & dosage; Phorbol Esters - chemistry; plasma membrane; Polyphenol; polyphenols; Presynaptic; Primary Cell Culture; Protein kinase C; Protein Kinase C-alpha - antagonists & inhibitors; Protein Kinase C-alpha - chemistry; Resveratrol; SNARE Proteins - chemistry; SNARE Proteins - metabolism; Stilbenes - administration & dosage; Synaptic Transmission - drug effects; Neuroprotection; Protein kinase C; BBB; PS; TPA; DMSO; HRP; PFA; Activation; EGFP; MTT; Membrane translocation; SNARE; DMEM; FITC; Polyphenol; FBS; MD; Resveratrol; FRET; Inhibition; Presynaptic; AD; DPPH; MALDI-TOF; GST; PKC; DAG; SDS-PAGE; Munc13-1; PEI; IPTG
• ISSN: 0304-4165; 0006-3002; 1872-8006; 1878-2434
• DOI: 10.1016/j.bbagen.2017.07.006

Resveratrol (1) is a naturally occurring polyphenol that has been implicated in neuroprotection. One of resveratrol's several biological targets is Ca2+-sensitive protein kinase C alpha (PKCα). Resveratrol inhibits PKCα by binding to its activator-binding C1 domain. Munc13-1 is a C1 domain-containing Ca2+-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 Munc13-1H567K, 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. [Display omitted] •Resveratrol binds to the C1 domain of SNARE complex protein Munc13-1•Resveratrol inhibits the activity of Munc13-1 in HT-22 cells and primary hippocampal neurons.•Dietary polyphenols are important in the management of neurodegenerative diseases.