Characterization of the furin cleavage motif for HIV-1 trimeric envelope glycoprotein by intact LC-MS analysis.
Analyst
; 145(5): 1636-1640, 2020 Mar 02.
Article
em En
| MEDLINE
| ID: mdl-31932825
ABSTRACT
Generating a soluble and native-like trimeric envelope glycoprotein (Env) with high efficacy as an immunogen has been a major focus for developing an effective vaccine against HIV-1. The Env immunogen is a heavily glycosylated protein composed of 3 identical surface gp120 and gp41 subunits that form into a trimer of heterodimers (3 × 28 N-glycan sites). During Env immunogen production, endogenous furin works to cleave a hexa-arginine motif connecting the gp120 and gp41 subunits, which is needed to ensure proper protein folding and a native-like conformation of Env. Verification of the overall identity and proteolytic cleavage of Env is therefore important for HIV-1 vaccine development and product quality. Herein, we report the first work using LC-MS to (1) achieve fast and accurate intact mass measurement of Env after deglycosylation and (2) confidently identify the furin cleavage sites.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteína gp41 do Envelope de HIV
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Proteína gp120 do Envelope de HIV
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Cromatografia Líquida
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Furina
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Espectrometria de Massas em Tandem
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Multimerização Proteica
Limite:
Humans
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article