title: SNARE zippering is hindered by polyphenols in the neuron.
abstract: Fusion of synaptic vesicles with the presynaptic plasma membrane in the neuron is mediated by soluble N-ethylmaleimide-sensitive fusion protein-attachment protein receptor (SNARE) proteins. SNARE complex formation is a zippering-like process which initiates at the N-terminus and proceeds to the C-terminal membrane-proximal region. Previously, we showed that this zippering-like process is regulated by several polyphenols, leading to the arrest of membrane fusion and the inhibition of neuroexocytosis. In vitro studies using purified SNARE proteins reconstituted in liposomes revealed that each polyphenol uniquely regulates SNARE zippering. However, the unique regulatory effect of each polyphenol in cells has not yet been examined. In the present study, we observed SNARE zippering in neuronal PC12 cells by measuring the fluorescence resonance energy transfer (FRET) changes of a cyan fluorescence protein (CFP) and a yellow fluorescence protein (YFP) fused to the N-termini or C-termini of SNARE proteins. We show that delphinidin and cyanidin inhibit the initial N-terminal nucleation of SNARE complex formation in a Ca(2+)-independent manner, while myricetin inhibits Ca(2+)-dependent transmembrane domain association of the SNARE complex in the cell. This result explains how polyphenols exhibit botulinum neurotoxin-like activity in vivo.
authors: Department of Genetic Engineering and Center for Human Interface Nanotechnology, Sungkyunkwan University, Suwon 440-746, South Korea; Biomedical Research Institute, Korea Institute of Science and Technology, Seoul 136-791, South Korea.
authors: Yoosoo
authors: Y
authors: Yang
authors: Department of Genetic Engineering and Center for Human Interface Nanotechnology, Sungkyunkwan University, Suwon 440-746, South Korea.
authors: Se-Hyun
authors: SH
authors: Kim
authors: Department of Genetic Engineering and Center for Human Interface Nanotechnology, Sungkyunkwan University, Suwon 440-746, South Korea.
authors: Paul
authors: P
authors: Heo
authors: Department of Genetic Engineering and Center for Human Interface Nanotechnology, Sungkyunkwan University, Suwon 440-746, South Korea.
authors: Byoungjae
authors: B
authors: Kong
authors: Department of Genetic Engineering and Center for Human Interface Nanotechnology, Sungkyunkwan University, Suwon 440-746, South Korea.
authors: Jonghyeok
authors: J
authors: Shin
authors: Department of Genetic Engineering and Center for Human Interface Nanotechnology, Sungkyunkwan University, Suwon 440-746, South Korea.
authors: Young-Hun
authors: YH
authors: Jung
authors: Department of Genetic Engineering and Center for Human Interface Nanotechnology, Sungkyunkwan University, Suwon 440-746, South Korea.
authors: Keejung
authors: K
authors: Yoon
authors: Department of Genetic Engineering and Center for Human Interface Nanotechnology, Sungkyunkwan University, Suwon 440-746, South Korea.
authors: Woo-Jae
authors: WJ
authors: Chung
authors: Biomedical Research Institute, Korea Institute of Science and Technology, Seoul 136-791, South Korea; Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, IA 50011, USA.
authors: Yeon-Kyun
authors: YK
authors: Shin
authors: Department of Genetic Engineering and Center for Human Interface Nanotechnology, Sungkyunkwan University, Suwon 440-746, South Korea. Electronic address: dhkweon@skku.edu.
authors: Dae-Hyuk
authors: DH
authors: Kweon
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D016350:Leucine Zippers / Q000187:drug effects*; D008561:Membrane Fusion / Q000187:drug effects / Q000502:physiology*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D016716:PC12 Cells; D059808:Polyphenols / Q000494:pharmacology*; D051381:Rats; D050600:SNARE Proteins / Q000378:metabolism*
chemical_list: D059808:Polyphenols; D050600:SNARE Proteins; D002118:Calcium
keywords: Botulinum toxin; FRET; Membrane fusion; Neurotransmitter; Polyphenol; SNARE