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TitleInfluences on trimerization and aggregation of soluble, cleaved HIV-1 SOSIP envelope glycoprotein.
Journal, issue, pagesJ Virol, Vol. 87, Issue 17, Page 9873-9885, Year 2013
Publish dateJul 3, 2013
AuthorsPer Johan Klasse / Rafael S Depetris / Robert Pejchal / Jean-Philippe Julien / Reza Khayat / Jeong Hyun Lee / Andre J Marozsan / Albert Cupo / Nicolette Cocco / Jacob Korzun / Anila Yasmeen / Andrew B Ward / Ian A Wilson / Rogier W Sanders / John P Moore /
PubMed AbstractWe describe methods to improve the properties of soluble, cleaved gp140 trimers of the human immunodeficiency virus type 1 (HIV-1) envelope glycoproteins (Env) for use in structural studies and as ...We describe methods to improve the properties of soluble, cleaved gp140 trimers of the human immunodeficiency virus type 1 (HIV-1) envelope glycoproteins (Env) for use in structural studies and as immunogens. In the absence of nonionic detergents, gp140 of the KNH1144 genotype, terminating at residue 681 in gp41 (SOSIP.681), has a tendency to form higher-order complexes or aggregates, which is particularly undesirable for structure-based research. We found that this aggregation in the absence of detergent does not involve the V1, V2, or V3 variable regions of gp120. Moreover, we observed that detergent forms micelles around the membrane-proximal external region (MPER) of the SOSIP.681 gp140 trimers, whereas deletion of most of the MPER residues by terminating the gp140 at residue 664 (SOSIP.664) prevented the aggregation that otherwise occurs in SOSIP.681 in the absence of detergent. Although the MPER can contribute to trimer formation, truncation of most of it only modestly reduced trimerization and lacked global adverse effects on antigenicity. Thus, the MPER deletion minimally influenced the kinetics of the binding of soluble CD4 and a CD4-binding site antibody to immobilized trimers, as detected by surface plasmon resonance. Furthermore, the MPER deletion did not alter the overall three-dimensional structure of the trimers, as viewed by negative-stain electron microscopy. Homogeneous and aggregate-free MPER-truncated SOSIP Env trimers are therefore useful for immunogenicity and structural studies.
External linksJ Virol / PubMed:23824824 / PubMed Central
MethodsEM (single particle)
Resolution14.0 - 22.0 Å
Structure data

EMDB-5700:
Negative stained image reconstruction of HIV KNH11444 subtype A SOSIP.681
Method: EM (single particle) / Resolution: 18.0 Å

EMDB-5701:
Negative stained image reconstruction of HIV KNH11444 subtype A SOSIP.681.dV1V2V3
Method: EM (single particle) / Resolution: 17.0 Å

EMDB-5702:
Negative stained image reconstruction of HIV KNH11444 subtype A SOSIP.681 in complex with Fab fragment of VRC PG04 monoclonal antibody
Method: EM (single particle) / Resolution: 15.0 Å

EMDB-5703:
Negative stained image reconstruction of HIV KNH11444 subtype A SOSIP.681.dV1V2V3 in complex with Fab fragment of VRC PG04 monoclonal antibody
Method: EM (single particle) / Resolution: 18.0 Å

EMDB-5704:
Negative stained image reconstruction of HIV KNH11444 subtype A SOSIP.681 in complex with soluble CD4
Method: EM (single particle) / Resolution: 22.0 Å

EMDB-5705:
Negative stained image reconstruction of HIV KNH11444 subtype A SOSIP.664
Method: EM (single particle) / Resolution: 14.0 Å

EMDB-5706:
Negative stained image reconstruction of HIV KNH11444 subtype A SOSIP.664 in complex with Fab fragment of VRC PG04 monoclonal antibody
Method: EM (single particle) / Resolution: 15.0 Å

EMDB-5707:
Negative stained image reconstruction of HIV KNH11444 subtype A SOSIP.664.dV1V2V3 in complex with Fab fragment of VRC PG04 monoclonal antibody
Method: EM (single particle) / Resolution: 19.0 Å

EMDB-5708:
Negative stained image reconstruction of HIV KNH11444 subtype A SOSIP.664 in complex with soluble CD4
Method: EM (single particle) / Resolution: 16.0 Å

EMDB-5709:
Negative stained image reconstruction of HIV KNH11444 subtype A SOSIP.664.dV1V2V3 in complex with soluble CD4
Method: EM (single particle) / Resolution: 19.0 Å

Source
  • Human immunodeficiency virus
  • unidentified (others)
  • Homo sapiens (human)

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