Fig. 1: Structure of OaVMAT2TM8/9-BRIL.
From: Engineering of a mammalian VMAT2 for cryo-EM analysis results in non-canonical protein folding
a Cryo-EM structure of OaVMAT2TM8/9-BRIL dimer. Left, OaVMAT2TM8/9-BRIL dimer viewed parallel to the membrane (gray rectangle). N- and C-domains of protomer A are colored in green and orange, respectively, while protomer B in light blue. BRIL modules are displayed as white surface. Right, OaVMAT2TM8/9-BRIL dimer viewed perpendicular to the membrane from the cytosolic side. Twelve TM helices of VMAT2 are labeled with numbers throughout the manuscript. BRIL modules are omitted for clearer views for the rest of the manuscript. b Superposition of OaVMAT2TM8/9-BRIL C-domain (in orange) onto OaVMAT2TM8/9-BRIL N-domain (in green). c Superposition of OaVMAT2TM8/9-BRIL protomer onto hMCT2 protomer (PDB: 7BP3) by aligning N-domains (green cartoon for OaVMAT2TM8/9-BRIL and pink cartoon for hMCT2). C-domains are displayed as orange surface for OaVMAT2TM8/9-BRIL and dark pink surface for hMCT2. d MST fitting curves of DA/SER binding to OaVMAT2 WT and variants as indicated, N = 3 repeats with biologically independent protein samples. Data are presented as mean ± SEM. Source data are provided as a Source Data file. e Docking of DA (magenta sticks) and SER (cyan sticks) in OaVMAT2TM8/9-BRIL. The cavity between N-domain (green cartoon) and C-domain (orange cartoon) is indicated by red mesh. f Potential polar interactions between docked DA (magenta sticks) or SER (cyan sticks) and OaVMAT2TM8/9-BRIL (green and orange cartoons) are indicated by black dashed lines. Participating residues are displayed as orange sticks. g Normalized FFN200 uptake by HEK293T cells expressing various VMAT2 WT and variants as indicated, N = 3 experiments with biologically independent cells cultured in different wells. Data are presented as mean ± SEM. Source data are provided as a Source Data file. Individual data points are shown as black dots. Two-tailed Student’s t-test was performed between the “TM8/9-BRIL” group and the “TM8/9-BRIL D402A” group (P = 0.012) for OaVMAT2, and between “WT” group and “E312A” group (P = 0.00022) or “D399A” group (P = 0.00049) for HsVMAT2. *P < 0.05; **P < 0.001.
