Characterization of Glycosylation Profiles of HIV-1 Transmitted/Founder Envelopes by Mass Spectrometry ▿ †
| Autor: | David Hua, Li Hua Ping, Eden P. Go, Heather Desaire, Hua-Xin Liao, Haiyan Chen, Jeffrey A. Anderson, Geetha S. Hewawasam, Barton F. Haynes |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Glycan
Glycosylation Glycoside Hydrolases medicine.medical_treatment Immunology HIV Infections Microbiology Mass Spectrometry law.invention chemistry.chemical_compound law Virology medicine Humans Trypsin Amino Acid Sequence chemistry.chemical_classification Protease biology HEK 293 cells env Gene Products Human Immunodeficiency Virus virus diseases Glycopeptide carbohydrates (lipids) Enzyme HEK293 Cells Biochemistry chemistry Insect Science biology.protein Recombinant DNA HIV-1 Pathogenesis and Immunity lipids (amino acids peptides and proteins) Antibody Sequence Alignment |
| Popis: | The analysis of HIV-1 envelope carbohydrates is critical to understanding their roles in HIV-1 transmission as well as in binding of envelope to HIV-1 antibodies. However, direct analysis of protein glycosylation by glycopeptide-based mass mapping approaches involves structural simplification of proteins with the use of a protease followed by an isolation and/or enrichment step before mass analysis. The successful completion of glycosylation analysis is still a major analytical challenge due to the complexity of samples, wide dynamic range of glycopeptide concentrations, and glycosylation heterogeneity. Here, we use a novel experimental workflow that includes an up-front complete or partial enzymatic deglycosylation step before trypsin digestion to characterize the glycosylation patterns and maximize the glycosylation coverage of two recombinant HIV-1 transmitted/founder envelope oligomers derived from clade B and C viruses isolated from acute infection and expressed in 293T cells. Our results show that both transmitted/founder Envs had similar degrees of glycosylation site occupancy as well as similar glycan profiles. Compared to 293T-derived recombinant Envs from viruses isolated from chronic HIV-1, transmitted/founder Envs displayed marked differences in their glycosylation site occupancies and in their amounts of complex glycans. Our analysis reveals that the glycosylation patterns of transmitted/founder Envs from two different clades (B and C) are more similar to each other than they are to the glycosylation patterns of chronic HIV-1 Envs derived from their own clades. |
| Databáze: | OpenAIRE |
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