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Characterization of Glycosylation Profiles of HIV-1 Transmitted/Founder Envelopes by Mass Spectrometry
Go, Eden P. ; Hewawasam, Geetha S. ; Lio, Hua-Xin ; Chen, Haiyan ; Ping, Li-Hua ; Anderson, Jeffrey A. ; Hua, David C. ; Haynes, Barton F. ; Desaire, Heather
Go, Eden P.
Hewawasam, Geetha S.
Lio, Hua-Xin
Chen, Haiyan
Ping, Li-Hua
Anderson, Jeffrey A.
Hua, David C.
Haynes, Barton F.
Desaire, Heather
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Abstract
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.
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This is the published version, also available here: http://dx.doi.org/10.1128/JVI.05053-11.
Date
2011-08-01
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American Society for Microbiology
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Go, Eden P., Hewawasam, Geetha., Lio, Hua-Xin., Chen, Haiyan., Ping, Li-Hua., Anderson, Jeffrey A., Hua, David C., Haynes, Barton F., Desaire, Heather. "Characterization of Glycosylation Profiles of HIV-1 Transmitted/Founder Envelopes by Mass Spectrometry." J. Virol. August 2011 vol. 85 no. 16 8270-8284. http://dx.doi.org/10.1128/JVI.05053-11.