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Here, we examined the physico-chemical requirements for membrane fusion of VLPs from West Nile and Japanese encephalitis viruses. In a bulk fusion assay, optimal hemifusion (or lipid mixing) efficiencies were observed at 37 \u00b0C. Fusion efficiency increased with decreasing pH; half-maximal hemifusion was attained at pH 5.6. The anionic lipids bis(monoacylglycero)phosphate and phosphatidylinositol-3-phosphate, when present in the target membrane, significantly enhanced fusion efficiency, consistent with the emerging model that flaviviruses fuse with intermediate-to-late endosomal compartments, where these lipids are most abundant. In a single-particle fusion assay, VLPs catalysed membrane hemifusion, tracked as lipid mixing with the cellular membrane, on a timescale of 7\u201320 s after acidification. Lipid mixing kinetics suggest that hemifusion is a kinetically complex, multistep process.<\/jats:p>","DOI":"10.1099\/vir.0.000113","type":"journal-article","created":{"date-parts":[[2015,3,5]],"date-time":"2015-03-05T02:24:19Z","timestamp":1425522259000},"page":"1702-1711","source":"Crossref","is-referenced-by-count":40,"title":["Physico-chemical requirements and kinetics of membrane fusion of flavivirus-like particles"],"prefix":"10.1099","volume":"96","author":[{"given":"Danillo L. 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