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. 2021 Feb 23;12:1238. doi: 10.1038/s41467-021-21505-9

Fig. 4. Formation of a lipid pocket by rearrangement of membrane-associated helices.

Fig. 4

a Position and organization of membrane-associated helices in immature E. Two amphipathic helices lie flat on the membrane (colored green and red), whereas two transmembrane (TM) helices span it (colored yellow and orange). b Close-up view of the membrane-associated helices of immature E and mature E of SPOV. Amphipathic helices are numbered H1 and H2, TM helices TM1 and TM2. In mature E (on the right), the amphipathic helices have reorganized, opening up a pocket that is filled with density resembling a phosphatidylethanolamine (PE) lipid. The lipid is shown in stick representation. The density fit is shown separately in c for clarity. Residues in close vicinity are shown as sticks and labeled appropriately. c Lipid fitted into the cryo-EM map at the binding pocket. d MSA of E sequences located next to the lipid, highlighting the conservation of H447 and G451 (SPOV numbering). Virus abbreviations as in Fig. 2. e Density present in the pocket of mature ZIKV (accession code: EMDB-7543). f Density present in the pocket of mature DENV2 (this study). g Structure of Sindbis virus E1 and E2 (PDB-ID: 6IMM) illustrating the similarity of the position of a glycoprotein associated lipid (colored in red). h Virus recovery after Gibson assembly of mutant viruses. Wild-type and mutants of DENV2/16681 were constructed via Gibson assembly and propagated in C6/36 cells. Production of virus was assessed after given intervals by ELISA. Cells infected with virus stock served as positive control. The results of n = 2 biologically independent experiments are shown.