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TitleCleavage-independent HIV-1 Env trimers engineered as soluble native spike mimetics for vaccine design.
Journal, issue, pagesCell Rep, Vol. 11, Issue 4, Page 539-550, Year 2015
Publish dateApr 28, 2015
AuthorsShailendra Kumar Sharma / Natalia de Val / Shridhar Bale / Javier Guenaga / Karen Tran / Yu Feng / Viktoriya Dubrovskaya / Andrew B Ward / Richard T Wyatt /
PubMed AbstractViral glycoproteins mediate entry by pH-activated or receptor-engaged activation and exist in metastable pre-fusogenic states that may be stabilized by directed rational design. As recently reported, ...Viral glycoproteins mediate entry by pH-activated or receptor-engaged activation and exist in metastable pre-fusogenic states that may be stabilized by directed rational design. As recently reported, the conformationally fixed HIV-1 envelope glycoprotein (Env) trimers in the pre-fusion state (SOSIP) display molecular homogeneity and structural integrity at relatively high levels of resolution. However, the SOSIPs necessitate full Env precursor cleavage, which requires endogenous furin overexpression. Here, we developed an alternative strategy using flexible peptide covalent linkage of Env subdomains to produce soluble, homogeneous, and cleavage-independent Env mimics, called native flexibly linked (NFL) trimers, as vaccine candidates. This simplified design avoids the need for furin co-expression and, in one case, antibody affinity purification to accelerate trimer scale-up for preclinical and clinical applications. We have successfully translated the NFL design to multiple HIV-1 subtypes, establishing the potential to become a general method of producing native-like, well-ordered Env trimers for HIV-1 or other viruses.
External linksCell Rep / PubMed:25892233 / PubMed Central
MethodsEM (single particle)
Resolution17.0 - 23.0 Å
Structure data

EMDB-6293:
JRFL gp140 NFL2P unliganded
Method: EM (single particle) / Resolution: 20.0 Å

EMDB-6294:
Bg505 gp140 NFL2P unliganded
Method: EM (single particle) / Resolution: 22.0 Å

EMDB-6295:
JRFL gp140 NFL2P liganded to PGV04
Method: EM (single particle) / Resolution: 22.0 Å

EMDB-6296:
Bg505 gp140 NFL2P liganded to PGV04
Method: EM (single particle) / Resolution: 17.0 Å

EMDB-6297:
JRFL gp140 NFL2 liganded to 2G12
Method: EM (single particle) / Resolution: 20.0 Å

EMDB-6298:
Bg505 gp140 NFL liganded to 2G12
Method: EM (single particle) / Resolution: 23.0 Å

Source
  • Human immunodeficiency virus 1
  • Homo sapiens (human)

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