Global site-specific N-glycosylation analysis of HIV envelope glycoprotein
| Autor: | Lin He, Matthias Pauthner, Yumiko Adachi, William R. Schief, James C. Paulson, Ching Yao Su, Dennis R. Burton, Erik Georgeson, Michael Kubitz, Claire M. Delahunty, Liwei Cao, John R. Yates, Raiees Andrabi, Javier Guenaga, Jolene K. Diedrich, Sung-Kyu Robin Park, Daniel W. Kulp, Sergey Menis, Devin Sok |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Models Molecular Glycan Glycosylation animal structures Science General Physics and Astronomy macromolecular substances Biology HIV Envelope Protein gp120 Proteomics General Biochemistry Genetics and Molecular Biology Article Mass Spectrometry law.invention 03 medical and health sciences chemistry.chemical_compound Epitopes N-linked glycosylation law Polysaccharides HIV vaccine chemistry.chemical_classification Multidisciplinary virus diseases Reproducibility of Results General Chemistry Virology 3. Good health carbohydrates (lipids) 030104 developmental biology chemistry biology.protein Recombinant DNA HIV-1 lipids (amino acids peptides and proteins) Antibody Protein Multimerization Glycoprotein Peptides |
| Zdroj: | Nature Communications Nature Communications, Vol 8, Iss 1, Pp 1-13 (2017) |
| ISSN: | 2041-1723 |
| Popis: | HIV-1 envelope glycoprotein (Env) is the sole target for broadly neutralizing antibodies (bnAbs) and the focus for design of an antibody-based HIV vaccine. The Env trimer is covered by ∼90N-linked glycans, which shield the underlying protein from immune surveillance. bNAbs to HIV develop during infection, with many showing dependence on glycans for binding to Env. The ability to routinely assess the glycan type at each glycosylation site may facilitate design of improved vaccine candidates. Here we present a general mass spectrometry-based proteomics strategy that uses specific endoglycosidases to introduce mass signatures that distinguish peptide glycosites that are unoccupied or occupied by high-mannose/hybrid or complex-type glycans. The method yields >95% sequence coverage for Env, provides semi-quantitative analysis of the glycosylation status at each glycosite. We find that most glycosites in recombinant Env trimers are fully occupied by glycans, varying in the proportion of high-mannose/hybrid and complex-type glycans. The analysis of site-specific glycosylation of HIV Envelope glycoprotein (Env) is challenging as it contains 25–30 glycosylation sites with multiple glycan forms at each site. Here the authors present a generally applicable mass spectrometry-based method for site-specific analysis of protein glycosylation that they apply to the analysis of the HIV-1 Env. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|