Yang, Yoosoo Kim, Se-Hyun Heo, Paul Kong, Byoungjae Shin, Jonghyeok Jung, Young-Hun Yoon, Keejung Chung, Woo-Jae Shin, Yeon-Kyun Kweon, Dae-Hyuk 2024-01-20T09:30:57Z 2024-01-20T09:30:57Z 2021-09-05 2014-07-18 0006-291X https://pubs.kist.re.kr/handle/201004/126586 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 Ca2+-independent manner, while myricetin inhibits Ca2+-dependent transmembrane domain association of the SNARE complex in the cell. This result explains how polyphenols exhibit botulinum neurotoxin-like activity in vivo. (C) 2014 Elsevier Inc. All rights reserved. English ACADEMIC PRESS INC ELSEVIER SCIENCE MEMBRANE-FUSION COMPLEX EXOCYTOSIS MECHANISM HEMIFUSION RELEASE CELLS NSF SNARE zippering is hindered by polyphenols in the neuron Article 10.1016/j.bbrc.2014.06.064 1 BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, v.450, no.1, pp.831 - 836 BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 450 1 831 836 scie scopus 000343641000137 2-s2.0-84907204799 Biochemistry & Molecular Biology Biophysics Biochemistry & Molecular Biology Biophysics Article MEMBRANE-FUSION COMPLEX EXOCYTOSIS MECHANISM HEMIFUSION RELEASE CELLS NSF SNARE Membrane fusion Botulinum toxin Neurotransmitter FRET Polyphenol