Structure-Based Design of Prefusion-Stabilized Filovirus Glycoprotein Trimers.
Amino Acid Substitution
Cell Line
Crystallography, X-Ray
Ebolavirus
/ metabolism
Filoviridae
/ metabolism
Glycoproteins
/ chemistry
Humans
Marburgvirus
/ metabolism
Models, Molecular
Mutation
/ genetics
Perfusion
Protein Domains
Protein Multimerization
Protein Stability
Structure-Activity Relationship
Ebola
Marburg
X-ray structure
crystal
filovirus
glycoprotein
instability
prefusion trimer
size-exclusion chromatography
stabilizing substitutions
Journal
Cell reports
ISSN: 2211-1247
Titre abrégé: Cell Rep
Pays: United States
ID NLM: 101573691
Informations de publication
Date de publication:
31 03 2020
31 03 2020
Historique:
received:
21
08
2019
revised:
27
01
2020
accepted:
10
03
2020
entrez:
3
4
2020
pubmed:
3
4
2020
medline:
27
3
2021
Statut:
ppublish
Résumé
Ebola virus causes severe hemorrhagic fever, often leading to death in humans. The trimeric fusion glycoprotein (GP) is the sole target for neutralizing antibodies and is the major focus of vaccine development. Soluble GP ectodomains are unstable and mostly monomeric when not fused to a heterologous trimerization domain. Here, we report structure-based designs of Ebola and Marburg GP trimers based on a stabilizing mutation in the hinge loop in refolding region 1 and substitution of a partially buried charge at the interface of the GP1 and GP2 subunits. The combined substitutions (T577P and K588F) substantially increased trimer expression for Ebola GP proteins. We determined the crystal structure of stabilized GP from the Makona Zaire ebolavirus strain without a trimerization domain or complexed ligand. The structure reveals that the stabilized GP adopts the same trimeric prefusion conformation, provides insight into triggering of GP conformational changes, and should inform future filovirus vaccine development.
Identifiants
pubmed: 32234486
pii: S2211-1247(20)30339-9
doi: 10.1016/j.celrep.2020.03.025
pmc: PMC7118701
pii:
doi:
Substances chimiques
Glycoproteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
4540-4550.e3Informations de copyright
Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Interests L.R., S.B., J.J., and J.P.M.L. are employees at Janssen Vaccines & Prevention. L.R., S.B., and J.P.M.L. are inventors on an international patent application describing trimer stabilizing Ebola GP mutations. J.P.M.L. holds stock in Johnson & Johnson. The remaining authors declare no competing financial interests.
Références
Proc Natl Acad Sci U S A. 2017 Aug 29;114(35):E7348-E7357
pubmed: 28807998
Acta Crystallogr D Biol Crystallogr. 2005 Jul;61(Pt 7):932-41
pubmed: 15983416
J Infect Dis. 2007 Nov 15;196 Suppl 2:S259-63
pubmed: 17940958
Nat Rev Microbiol. 2019 May;17(5):261-263
pubmed: 30926957
Proc Natl Acad Sci U S A. 2018 Nov 27;115(48):12265-12270
pubmed: 30420505
Virology. 2000 Dec 5;278(1):20-6
pubmed: 11112476
Nature. 2008 Jul 10;454(7201):177-82
pubmed: 18615077
Science. 2017 Jun 2;356(6341):923-928
pubmed: 28572385
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
pubmed: 15572765
Acta Crystallogr D Biol Crystallogr. 2002 Nov;58(Pt 11):1948-54
pubmed: 12393927
Nat Commun. 2015 Sep 03;6:8143
pubmed: 26333350
PLoS One. 2019 Jul 5;14(7):e0219312
pubmed: 31276481
Virology. 2011 Jul 5;415(2):83-94
pubmed: 21529875
Cell Rep. 2018 Sep 4;24(10):2723-2732.e4
pubmed: 30184505
Nat Commun. 2018 Sep 26;9(1):3934
pubmed: 30258051
EMBO J. 2012 Apr 18;31(8):1947-60
pubmed: 22395071
Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3
pubmed: 15299374
Nature. 2011 Aug 24;477(7364):340-3
pubmed: 21866103
Proc Natl Acad Sci U S A. 2011 May 17;108(20):8426-31
pubmed: 21536871
Nat Commun. 2017 Nov 16;8(1):1528
pubmed: 29142300
mBio. 2016 Feb 23;7(1):e02154-15
pubmed: 26908579
PLoS Pathog. 2013 Mar;9(3):e1003232
pubmed: 23555248
PLoS Pathog. 2010 Sep 23;6(9):e1001121
pubmed: 20886108
Nature. 2011 Aug 24;477(7364):344-8
pubmed: 21866101
Cell. 2016 Jan 14;164(1-2):258-268
pubmed: 26771495
J Virol. 2002 Sep;76(17):8875-89
pubmed: 12163607
Nat Protoc. 2017 Jul;12(7):1310-1325
pubmed: 28569763
mBio. 2018 Sep 11;9(5):
pubmed: 30206174
Immunity. 2017 May 16;46(5):792-803.e3
pubmed: 28514686
Cell Rep. 2018 Apr 10;23(2):584-595
pubmed: 29642014
Virology. 2011 Oct 25;419(2):72-83
pubmed: 21907381
J Virol. 2000 Jan;74(2):627-43
pubmed: 10623724
J Virol. 2006 Apr;80(8):4174-8
pubmed: 16571833
Science. 2005 Jun 10;308(5728):1643-5
pubmed: 15831716
Acta Crystallogr D Biol Crystallogr. 2003 Jul;59(Pt 7):1131-7
pubmed: 12832755
Nat Struct Mol Biol. 2011 Nov 20;18(12):1424-7
pubmed: 22101933
Virology. 2003 Jan 5;305(1):115-23
pubmed: 12504546
Acta Crystallogr D Biol Crystallogr. 2013 Jul;69(Pt 7):1204-14
pubmed: 23793146
Elife. 2013 Sep 10;2:e01456
pubmed: 24040512
Viruses. 2012 Apr;4(4):447-70
pubmed: 22590681
PLoS Pathog. 2010 Sep 16;6(9):e1001110
pubmed: 20862315
J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674
pubmed: 19461840
Nature. 2016 Jul 7;535(7610):169-172
pubmed: 27362232
Science. 2016 Mar 18;351(6279):1339-42
pubmed: 26917593
J Virol. 2002 Jul;76(13):6841-4
pubmed: 12050398
Science. 2013 Nov 1;342(6158):592-8
pubmed: 24179220
Nat Microbiol. 2016 Aug 08;1(9):16128
pubmed: 27562261
mBio. 2019 Jul 9;10(4):
pubmed: 31289183