Influences on trimerization and aggregation of soluble, cleaved HIV-1 SOSIP envelope glycoprotein.

TitleInfluences on trimerization and aggregation of soluble, cleaved HIV-1 SOSIP envelope glycoprotein.
Publication TypeJournal Article
Year of Publication2013
AuthorsKlasse P J, Depetris RS, Pejchal R, Julien J-P, Khayat R, Lee J H, Marozsan AJ, Cupo A, Cocco N, Korzun J, Yasmeen A, Ward AB, Wilson IA, Sanders RW, Moore JP
JournalJ Virol
Volume87
Issue17
Pagination9873-85
Date Published09/01/2013
ISSN1098-5514
KeywordsAmino Acid Sequence, Antibodies, Monoclonal, Antibodies, Neutralizing, Antigens, CD4, Binding Sites, Detergents, env Gene Products, Human Immunodeficiency Virus, HEK293 Cells, HIV Antibodies, HIV Envelope Protein gp120, HIV Envelope Protein gp41, HIV-1, Humans, Microscopy, Electron, Molecular Sequence Data, Protein Multimerization, Protein Structure, Quaternary, Recombinant Proteins, Sequence Deletion, Solubility
Abstract

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.

DOI10.1128/JVI.01226-13
Alternate JournalJ. Virol.
PubMed ID23824824
PubMed Central IDPMC3754145
Grant ListP01 AI82362 / AI / NIAID NIH HHS / United States
P30 AI036214 / AI / NIAID NIH HHS / United States
P41 GM103310 / GM / NIGMS NIH HHS / United States
RR017573 / RR / NCRR NIH HHS / United States
UM1 AI100663 / AI / NIAID NIH HHS / United States
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