Kimmo Rantalainen / Zachary T Berndsen / Aleksandar Antanasijevic / Torben Schiffner / Xi Zhang / Wen-Hsin Lee / Jonathan L Torres / Lei Zhang / Adriana Irimia / Jeffrey Copps / Kenneth H Zhou / Young D Kwon / William H Law / Chaim A Schramm / Raffaello Verardi / Shelly J Krebs / Peter D Kwong / Nicole A Doria-Rose / Ian A Wilson / Michael B Zwick / John R Yates / William R Schief / Andrew B Ward /
PubMed 要旨
Structural and functional studies of HIV envelope glycoprotein (Env) as a transmembrane protein have long been complicated by challenges associated with inherent flexibility of the molecule and the ...Structural and functional studies of HIV envelope glycoprotein (Env) as a transmembrane protein have long been complicated by challenges associated with inherent flexibility of the molecule and the membrane-embedded hydrophobic regions. Here, we present approaches for incorporating full-length, wild-type HIV-1 Env, as well as C-terminally truncated and stabilized versions, into lipid assemblies, providing a modular platform for Env structural studies by single particle electron microscopy. We reconstitute a full-length Env clone into a nanodisc, complex it with a membrane-proximal external region (MPER) targeting antibody 10E8, and structurally define the full quaternary epitope of 10E8 consisting of lipid, MPER, and ectodomain contacts. By aligning this and other Env-MPER antibody complex reconstructions with the lipid bilayer, we observe evidence of Env tilting as part of the neutralization mechanism for MPER-targeting antibodies. We also adapt the platform toward vaccine design purposes by introducing stabilizing mutations that allow purification of unliganded Env with a peptidisc scaffold.
EMDB-21321: Full-length HIV-1 Envelope glycoprotein clone PC64 in complex with PGT151 Fab and VRC42.01 Fab 手法: EM (単粒子) / 解像度: 7.4 Å
EMDB-21322: Full-length HIV-1 Envelope glycoprotein clone AMC011 in complex with PGT151 Fab and VRC42.01 Fab 手法: EM (単粒子) / 解像度: 7.2 Å
EMDB-21323: Full-length HIV-1 Envelope glycoprotein clone PC64 in complex with PGT151 Fab and VRC42.N1 Fab 手法: EM (単粒子) / 解像度: 8.6 Å
EMDB-21324: Full-length HIV-1 Envelope glycoprotein clone PC64 in complex with PGT151 Fab and DH511.2 Fab 手法: EM (単粒子) / 解像度: 7.6 Å
EMDB-21326: Full-length HIV-1 Envelope glycoprotein clone PC64FL in complex with PGT151 Fab and VRC46.01 Fab 手法: EM (単粒子) / 解像度: 9.0 Å
EMDB-21327: Full-length HIV-1 Envelope glycoprotein clone AMC011 in complex with PGT151 Fab and two copies of PGZL1 Fab 手法: EM (単粒子) / 解像度: 6.6 Å
EMDB-21328: Full-length HIV-1 Envelope glycoprotein clone AMC011 in complex with PGT151 Fab and 10E8v4-5R+100cF Fab 手法: EM (単粒子) / 解像度: 7.8 Å
EMDB-21329: Nanodisc of C-terminally truncated HIV-1 Envelope glycoprotein clone BG505 in complex with PGT151 Fab 手法: EM (単粒子) / 解像度: 10.0 Å
EMDB-21330: Ectodomain from nanodisc of C-terminally truncated HIV-1 Envelope glycoprotein clone BG505 in complex with PGT151 Fab 手法: EM (単粒子) / 解像度: 4.6 Å
EMDB-21331: Nanodisc of full-length HIV-1 Envelope glycoprotein clone PC64FL in complex with PGT151 Fab 手法: EM (単粒子) / 解像度: 9.1 Å
EMDB-21332: Nanodisc of full-length HIV-1 Envelope glycoprotein clone AMC011 in complex with two PGT151 Fabs and one 10E8 Fab 手法: EM (単粒子) / 解像度: 6.5 Å
EMDB-21333: Nanodisc of full-length HIV-1 Envelope glycoprotein clone AMC011 in complex with two PGT151 Fabs and three 10E8 Fabs 手法: EM (単粒子) / 解像度: 9.4 Å
EMDB-21334: Nanodisc of full-length HIV-1 Envelope glycoprotein clone AMC011 in complex with one PGT151 Fab and two 10E8 Fabs 手法: EM (単粒子) / 解像度: 8.2 Å
EMDB-21335, PDB-6vpx: Nanodisc of full-length HIV-1 Envelope glycoprotein clone AMC011 in complex with one PGT151 Fab and three 10E8 Fabs 手法: EM (単粒子) / 解像度: 5.0 Å
EMDB-21336: Full-length HIV-1 Envelope glycoprotein clone PC64 in DOPC-CHS bicelle 手法: EM (単粒子) / 解像度: 19.0 Å
EMDB-21337: Complex of full-length HIV-1 Envelope glycoprotein clone PC64 and VRC42.01 Fab in DOPC-DOPS-CHS bicelle 手法: EM (単粒子) / 解像度: 18.0 Å
EMDB-21338: Complex of full-length HIV-1 Envelope glycoprotein clone PC64 and DH511.2 Fab in DOPC-DOPS-CHS bicelle 手法: EM (単粒子) / 解像度: 20.0 Å
EMDB-21339: Complex of full-length HIV-1 Envelope glycoprotein clone PC64 and PGT151 Fab in DOPC-DOPS nanodisc 手法: EM (単粒子) / 解像度: 18.0 Å
EMDB-21340: Complex of full-length HIV-1 Envelope glycoprotein clone PC64 and VRC42.N1 Fab in DOPC-DOPS-CHS bicelle 手法: EM (単粒子) / 解像度: 17.0 Å
EMDB-21341: Complex of full-length HIV-1 Envelope glycoprotein clone AMC011 and VRC42.01 Fab in DOPC-DOPS-CHS nanodisc 手法: EM (単粒子) / 解像度: 19.0 Å
EMDB-21342: Complex of full-length HIV-1 Envelope glycoprotein clone AMC011 and 10E8 Fab in nanodisc 手法: EM (単粒子) / 解像度: 26.0 Å
EMDB-21343: HIV-1 Envelope glycoprotein clone BG505-ST-710 in DOPC-DOPS-CHS peptidisc 手法: EM (単粒子) / 解像度: 18.0 Å
EMDB-21344: Complex of HIV-1 Envelope glycoprotein clone BG505-ST-710 and 10E8 Fab in DOPC-DOPS-CHS peptidisc 手法: EM (単粒子) / 解像度: 20.0 Å