Visualization of the HIV-1 Env glycan shield across scales.
Antibodies, Neutralizing
/ immunology
Antibody Formation
Computer Simulation
Cryoelectron Microscopy
/ methods
Epitopes
/ chemistry
Glycosylation
HIV Antibodies
/ immunology
HIV Envelope Protein gp120
/ metabolism
HIV Seropositivity
HIV-1
/ immunology
Humans
Immune Evasion
/ immunology
Mass Spectrometry
/ methods
Models, Molecular
Polysaccharides
/ metabolism
env Gene Products, Human Immunodeficiency Virus
/ chemistry
HIV-1
cryo-EM
glycoprotein
molecular modeling
vaccine
Journal
Proceedings of the National Academy of Sciences of the United States of America
ISSN: 1091-6490
Titre abrégé: Proc Natl Acad Sci U S A
Pays: United States
ID NLM: 7505876
Informations de publication
Date de publication:
10 11 2020
10 11 2020
Historique:
pubmed:
24
10
2020
medline:
8
1
2021
entrez:
23
10
2020
Statut:
ppublish
Résumé
The dense array of N-linked glycans on the HIV-1 envelope glycoprotein (Env), known as the "glycan shield," is a key determinant of immunogenicity, yet intrinsic heterogeneity confounds typical structure-function analysis. Here, we present an integrated approach of single-particle electron cryomicroscopy (cryo-EM), computational modeling, and site-specific mass spectrometry (MS) to probe glycan shield structure and behavior at multiple levels. We found that dynamics lead to an extensive network of interglycan interactions that drive the formation of higher-order structure within the glycan shield. This structure defines diffuse boundaries between buried and exposed protein surface and creates a mapping of potentially immunogenic sites on Env. Analysis of Env expressed in different cell lines revealed how cryo-EM can detect subtle changes in glycan occupancy, composition, and dynamics that impact glycan shield structure and epitope accessibility. Importantly, this identified unforeseen changes in the glycan shield of Env obtained from expression in the same cell line used for vaccine production. Finally, by capturing the enzymatic deglycosylation of Env in a time-resolved manner, we found that highly connected glycan clusters are resistant to digestion and help stabilize the prefusion trimer, suggesting the glycan shield may function beyond immune evasion.
Identifiants
pubmed: 33093196
pii: 2000260117
doi: 10.1073/pnas.2000260117
pmc: PMC7668054
doi:
Substances chimiques
Antibodies, Neutralizing
0
Epitopes
0
HIV Antibodies
0
HIV Envelope Protein gp120
0
Polysaccharides
0
env Gene Products, Human Immunodeficiency Virus
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
28014-28025Subventions
Organisme : NIGMS NIH HHS
ID : P41 GM103533
Pays : United States
Organisme : NIAID NIH HHS
ID : UM1 AI144462
Pays : United States
Organisme : NIAID NIH HHS
ID : UM1 AI100645
Pays : United States
Organisme : NIAID NIH HHS
ID : UM1 AI100663
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI113867
Pays : United States
Organisme : NIAID NIH HHS
ID : R56 AI113867
Pays : United States
Organisme : NIAID NIH HHS
ID : F31 AI131873
Pays : United States
Organisme : NIAID NIH HHS
ID : UM1 AI144371
Pays : United States
Informations de copyright
Copyright © 2020 the Author(s). Published by PNAS.
Déclaration de conflit d'intérêts
The authors declare no competing interest.
Références
Elife. 2016 Nov 15;5:
pubmed: 27845625
Sci Rep. 2017 Jun 30;7(1):4435
pubmed: 28667249
Nat Commun. 2017 Mar 28;8:14954
pubmed: 28348411
Structure. 2015 Oct 6;23(10):1943-1951
pubmed: 26388028
Nature. 1998 Jun 18;393(6686):648-59
pubmed: 9641677
Proc Natl Acad Sci U S A. 2008 Jun 17;105(24):8256-61
pubmed: 18550810
Science. 2019 Dec 6;366(6470):
pubmed: 31672916
Cell Rep. 2016 Mar 22;14(11):2695-706
pubmed: 26972002
J Phys Chem B. 2012 Jan 12;116(1):390-400
pubmed: 22118044
J Virol. 2004 Nov;78(22):12625-37
pubmed: 15507649
Proc Natl Acad Sci U S A. 2010 Aug 3;107(31):13800-5
pubmed: 20643940
Acta Crystallogr D Struct Biol. 2018 Apr 1;74(Pt 4):256-263
pubmed: 29652253
J Biol Chem. 1990 Jun 25;265(18):10373-82
pubmed: 2355006
Cell Mol Life Sci. 2007 Aug;64(16):2133-52
pubmed: 17558468
Cell. 2015 Apr 23;161(3):438-449
pubmed: 25910204
Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3131-6
pubmed: 19204290
J Comput Chem. 2004 Oct;25(13):1605-12
pubmed: 15264254
Glycobiology. 2015 Dec;25(12):1323-4
pubmed: 26543186
Nature. 2016 May 04;533(7604):561-4
pubmed: 27144356
J Virol. 1998 Oct;72(10):8365-70
pubmed: 9733886
Nat Methods. 2017 Mar;14(3):290-296
pubmed: 28165473
Elife. 2016 Sep 26;5:
pubmed: 27669148
PLoS Pathog. 2017 Sep 13;13(9):e1006614
pubmed: 28902916
Glycobiology. 2015 Dec;25(12):1335-49
pubmed: 26240167
Curr Opin Struct Biol. 2009 Oct;19(5):524-33
pubmed: 19647993
Immunity. 2019 Nov 19;51(5):915-929.e7
pubmed: 31732167
Curr Opin Struct Biol. 2019 Jun;56:37-45
pubmed: 30502729
Biochemistry. 1989 Jun 27;28(13):5536-43
pubmed: 2775722
Eur J Biochem. 1993 Nov 15;218(1):239-44
pubmed: 8243469
Sci Rep. 2018 Oct 9;8(1):15031
pubmed: 30302011
Science. 2013 Dec 20;342(6165):1484-90
pubmed: 24179160
PLoS Pathog. 2020 Aug 31;16(8):e1008753
pubmed: 32866207
Nat Commun. 2018 Sep 12;9(1):3693
pubmed: 30209313
Immunol Rev. 2017 Jan;275(1):161-182
pubmed: 28133806
Cell. 2016 May 5;165(4):813-26
pubmed: 27114034
Sci Rep. 2015 Mar 09;5:8926
pubmed: 25748215
Nature. 2017 Jul 20;547(7663):360-363
pubmed: 28700571
J Proteome Res. 2018 Mar 2;17(3):987-999
pubmed: 29420040
Nat Commun. 2015 Jun 24;6:7479
pubmed: 26105115
J Virol. 2016 Sep 29;90(20):9224-36
pubmed: 27489265
Science. 2016 Mar 4;351(6277):1043-8
pubmed: 26941313
Science. 2001 Mar 23;291(5512):2364-9
pubmed: 11269317
Virology. 2004 Dec 5;330(1):233-48
pubmed: 15527849
PLoS Pathog. 2013 Sep;9(9):e1003618
pubmed: 24068931
J Biol Chem. 2012 Jul 13;287(29):24239-54
pubmed: 22645128
Curr Opin Chem Biol. 1999 Dec;3(6):643-9
pubmed: 10600722
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
pubmed: 15572765
Biophys J. 2018 Apr 10;114(7):1604-1613
pubmed: 29642030
PLoS Comput Biol. 2016 Oct 7;12(10):e1005094
pubmed: 27716795
Biotechnol Bioeng. 2018 Apr;115(4):885-899
pubmed: 29150937
J Struct Biol. 2007 Jan;157(1):47-55
pubmed: 16931051
Elife. 2018 Nov 09;7:
pubmed: 30412051
Nature. 2003 Mar 20;422(6929):307-12
pubmed: 12646921
Science. 1986 Jul 11;233(4760):209-12
pubmed: 3014647
PLoS One. 2013 Nov 26;8(11):e80301
pubmed: 24303005
Mol Med Today. 1995 Sep;1(6):270-7
pubmed: 9415161
Sci Rep. 2018 Oct 30;8(1):16017
pubmed: 30375453
Retrovirology. 2013 Feb 06;10:14
pubmed: 23384254
Curr Opin Struct Biol. 2011 Oct;21(5):576-82
pubmed: 21978957
Cell Rep. 2017 Apr 25;19(4):719-732
pubmed: 28445724
Nat Struct Biol. 1996 Jan;3(1):45-53
pubmed: 8548454
Proc Natl Acad Sci U S A. 2012 Mar 27;109(13):4875-80
pubmed: 22403063
Structure. 2019 Jan 2;27(1):175-188.e6
pubmed: 30393052
PLoS One. 2018 Jan 12;13(1):e0191157
pubmed: 29329315
Immunity. 2018 Aug 21;49(2):288-300.e8
pubmed: 30097292
Cell Rep. 2020 Mar 17;30(11):3755-3765.e7
pubmed: 32187547
Immunol Rev. 2017 Jan;275(1):21-32
pubmed: 28133813
Br Med Bull. 2001;58:19-42
pubmed: 11714622
Trends Biochem Sci. 1994 Feb;19(2):65-70
pubmed: 8160267
J Biol Chem. 2017 Jun 16;292(24):10197-10219
pubmed: 28446609
Structure. 2017 Oct 3;25(10):1631-1639.e2
pubmed: 28890362
Structure. 2013 Mar 5;21(3):321-31
pubmed: 23473666