Lack of complex N-glycans on HIV-1 envelope glycoproteins preserves protein conformation and entry function

Dirk Eggink; Mark Melchers; Manfred Wuhrer; Thijs van Montfort; Antu K. dey; Benno A. Naaijkens; Kathryn B. David; Valentin Le Douce; André M. Deelder; Kenneth Kang; William C. Olson; Ben Berkhout; Cornelis H. Hokke; John P. Moore; Rogier W. Sanders

doi:https://doi.org/10.1016/j.virol.2010.02.019

Lack of complex N-glycans on HIV-1 envelope glycoproteins preserves protein conformation and entry function

Dirk Eggink, Mark Melchers, Manfred Wuhrer, Thijs van Montfort, Antu K. dey, Benno A. Naaijkens, Kathryn B. David, Valentin Le Douce, André M. Deelder, Kenneth Kang, William C. Olson, Ben Berkhout, Cornelis H. Hokke, John P. Moore, Rogier W. Sanders

Research output: Contribution to journal › Article › Academic › peer-review

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Abstract

The HIV-1 envelope glycoprotein complex (Env) is the focus of vaccine development aimed at eliciting humoral immunity. Env's extensive and heterogeneous N-linked glycosylation affects folding, binding to lectin receptors, antigenicity and immunogenicity. We characterized recombinant Env proteins and virus particles produced in mammalian cells that lack N-acetylglucosaminyltransferase I (GnTI), an enzyme necessary for the conversion of oligomannose N-glycans to complex N-glycans. Carbohydrate analyses revealed that trimeric Env produced in GnTI(-/-) cells contained exclusively oligomannose N-glycans, with incompletely trimmed oligomannose glycans predominating. The folding and conformation of Env proteins was little affected by the manipulation of the glycosylation. Viruses produced in GnTI(-/-) cells were infectious, indicating that the conversion to complex glycans is not necessary for Env entry function, although virus binding to the C-type lectin DC-SIGN was enhanced. Manipulating Env's N-glycosylation may be useful for structural and functional studies and for vaccine design. (C) 2010 Elsevier Inc. All rights reserved

Original language	English
Pages (from-to)	236-247
Journal	Virology
Volume	401
Issue number	2
DOIs	https://doi.org/10.1016/j.virol.2010.02.019
Publication status	Published - 2010

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https://doi.org/10.1016/j.virol.2010.02.019

Cite this

Eggink, D., Melchers, M., Wuhrer, M., van Montfort, T., dey, A. K., Naaijkens, B. A., David, K. B., Le Douce, V., Deelder, A. M., Kang, K., Olson, W. C., Berkhout, B., Hokke, C. H., Moore, J. P., & Sanders, R. W. (2010). Lack of complex N-glycans on HIV-1 envelope glycoproteins preserves protein conformation and entry function. Virology, 401(2), 236-247. https://doi.org/10.1016/j.virol.2010.02.019

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title = "Lack of complex N-glycans on HIV-1 envelope glycoproteins preserves protein conformation and entry function",

abstract = "The HIV-1 envelope glycoprotein complex (Env) is the focus of vaccine development aimed at eliciting humoral immunity. Env's extensive and heterogeneous N-linked glycosylation affects folding, binding to lectin receptors, antigenicity and immunogenicity. We characterized recombinant Env proteins and virus particles produced in mammalian cells that lack N-acetylglucosaminyltransferase I (GnTI), an enzyme necessary for the conversion of oligomannose N-glycans to complex N-glycans. Carbohydrate analyses revealed that trimeric Env produced in GnTI(-/-) cells contained exclusively oligomannose N-glycans, with incompletely trimmed oligomannose glycans predominating. The folding and conformation of Env proteins was little affected by the manipulation of the glycosylation. Viruses produced in GnTI(-/-) cells were infectious, indicating that the conversion to complex glycans is not necessary for Env entry function, although virus binding to the C-type lectin DC-SIGN was enhanced. Manipulating Env's N-glycosylation may be useful for structural and functional studies and for vaccine design. (C) 2010 Elsevier Inc. All rights reserved",

author = "Dirk Eggink and Mark Melchers and Manfred Wuhrer and {van Montfort}, Thijs and dey, {Antu K.} and Naaijkens, {Benno A.} and David, {Kathryn B.} and {Le Douce}, Valentin and Deelder, {Andr{\'e} M.} and Kenneth Kang and Olson, {William C.} and Ben Berkhout and Hokke, {Cornelis H.} and Moore, {John P.} and Sanders, {Rogier W.}",

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Eggink, D, Melchers, M, Wuhrer, M, van Montfort, T, dey, AK, Naaijkens, BA, David, KB, Le Douce, V, Deelder, AM, Kang, K, Olson, WC, Berkhout, B, Hokke, CH, Moore, JP & Sanders, RW 2010, 'Lack of complex N-glycans on HIV-1 envelope glycoproteins preserves protein conformation and entry function', Virology, vol. 401, no. 2, pp. 236-247. https://doi.org/10.1016/j.virol.2010.02.019

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AU - Naaijkens, Benno A.

AU - David, Kathryn B.

AU - Le Douce, Valentin

AU - Deelder, André M.

AU - Kang, Kenneth

AU - Olson, William C.

AU - Berkhout, Ben

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AB - The HIV-1 envelope glycoprotein complex (Env) is the focus of vaccine development aimed at eliciting humoral immunity. Env's extensive and heterogeneous N-linked glycosylation affects folding, binding to lectin receptors, antigenicity and immunogenicity. We characterized recombinant Env proteins and virus particles produced in mammalian cells that lack N-acetylglucosaminyltransferase I (GnTI), an enzyme necessary for the conversion of oligomannose N-glycans to complex N-glycans. Carbohydrate analyses revealed that trimeric Env produced in GnTI(-/-) cells contained exclusively oligomannose N-glycans, with incompletely trimmed oligomannose glycans predominating. The folding and conformation of Env proteins was little affected by the manipulation of the glycosylation. Viruses produced in GnTI(-/-) cells were infectious, indicating that the conversion to complex glycans is not necessary for Env entry function, although virus binding to the C-type lectin DC-SIGN was enhanced. Manipulating Env's N-glycosylation may be useful for structural and functional studies and for vaccine design. (C) 2010 Elsevier Inc. All rights reserved

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