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    Characterization of Glycosylation Profiles of HIV-1 Transmitted/Founder Envelopes by Mass Spectrometry

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    DesaireH_Aug2011.pdf (4.560Mb)
    Issue Date
    2011-08-01
    Author
    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
    Publisher
    American Society for Microbiology
    Type
    Article
    Article Version
    Scholarly/refereed, publisher version
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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.
    Description
    This is the published version, also available here: http://dx.doi.org/10.1128/JVI.05053-11.
    URI
    http://hdl.handle.net/1808/16043
    DOI
    https://doi.org/10.1128/JVI.05053-11
    Collections
    • Chemistry Scholarly Works [626]
    Citation
    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.

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    Contact KU ScholarWorks
    785-864-8983
    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    785-864-8983

    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    Image Credits
     

     

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