Characterization of glycosylation profiles of HIV-1 transmitted/founder envelopes by mass spectrometry.
Journal Article
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.
Full Text
Duke Authors
Cited Authors
- Go, EP; Hewawasam, G; Liao, H-X; Chen, H; Ping, L-H; Anderson, JA; Hua, DC; Haynes, BF; Desaire, H
Published Date
- August 2011
Published In
Volume / Issue
- 85 / 16
Start / End Page
- 8270 - 8284
PubMed ID
- 21653661
Pubmed Central ID
- 21653661
Electronic International Standard Serial Number (EISSN)
- 1098-5514
Digital Object Identifier (DOI)
- 10.1128/JVI.05053-11
Language
- eng
Conference Location
- United States