Identification of N-glycans with GalNAc-containing antennae from recombinant HIV trimers by ion mobility and negative ion fragmentation
Identification of N-glycans with GalNAc-containing antennae from recombinant HIV trimers by ion mobility and negative ion fragmentation
Negative ion collision-induced dissociation (CID) of underivatized N-glycans has proved to be a simple, yet powerful method for their structural determination. Recently, we have identified a series of such structures with GalNAc rather than the more common galactose capping the antennae of hybrid and complex glycans. As part of a series of publications describing the negative ion fragmentation of different types of N-glycan, this paper describes their CID spectra and estimated nitrogen cross sections recorded by travelling wave ion mobility mass spectrometry (TWIMS). Most of the glycans were derived from the recombinant glycoproteins gp120 and gp41 from the human immunodeficiency virus (HIV), recombinantly derived from human embryonic kidney (HEK 293T) cells. Twenty-six GalNAc-capped hybrid and complex N-glycans were identified by a combination of TWIMS, negative ion CID, and exoglycosidase digestions. They were present as the neutral glycans and their sulfated and α2→3-linked sialylated analogues. Overall, negative ion fragmentation of glycans generates fingerprints that reveal their structural identity.
Human immunodeficiency virus, Ion mobility, N-Acetylgalactosamine, N-Glycans, Negative ion fragmentation
7229-7240
Harvey, David J.
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Behrens, Anna-Janina
ed584c40-79cb-4de9-bb9c-bd68c71d6a68
Crispin, Max
cd980957-0943-4b89-b2b2-710f01f33bc9
Struwe, Weston B.
16a348b1-3921-4a2d-b5fb-d341fccea65f
December 2021
Harvey, David J.
8bb24417-3852-4b1f-827b-0d5d2c176744
Behrens, Anna-Janina
ed584c40-79cb-4de9-bb9c-bd68c71d6a68
Crispin, Max
cd980957-0943-4b89-b2b2-710f01f33bc9
Struwe, Weston B.
16a348b1-3921-4a2d-b5fb-d341fccea65f
Harvey, David J., Behrens, Anna-Janina, Crispin, Max and Struwe, Weston B.
(2021)
Identification of N-glycans with GalNAc-containing antennae from recombinant HIV trimers by ion mobility and negative ion fragmentation.
Analytical and Bioanalytical Chemistry, 413 (29), .
(doi:10.1007/s00216-021-03477-3).
Abstract
Negative ion collision-induced dissociation (CID) of underivatized N-glycans has proved to be a simple, yet powerful method for their structural determination. Recently, we have identified a series of such structures with GalNAc rather than the more common galactose capping the antennae of hybrid and complex glycans. As part of a series of publications describing the negative ion fragmentation of different types of N-glycan, this paper describes their CID spectra and estimated nitrogen cross sections recorded by travelling wave ion mobility mass spectrometry (TWIMS). Most of the glycans were derived from the recombinant glycoproteins gp120 and gp41 from the human immunodeficiency virus (HIV), recombinantly derived from human embryonic kidney (HEK 293T) cells. Twenty-six GalNAc-capped hybrid and complex N-glycans were identified by a combination of TWIMS, negative ion CID, and exoglycosidase digestions. They were present as the neutral glycans and their sulfated and α2→3-linked sialylated analogues. Overall, negative ion fragmentation of glycans generates fingerprints that reveal their structural identity.
Text
Harvey_AnalBioanalChem_2021_Article_IdentificationOfN-glycansWithG
More information
Accepted/In Press date: 2021
e-pub ahead of print date: 30 July 2021
Published date: December 2021
Additional Information:
Funding Information:
The work was supported by a the International AIDS Vaccine Initiative (IAVI) through grant INV-008352/OPP1153692 and the IAVI Neutralizing Antibody Center through the Collaboration for AIDS Vaccine Discovery grant OPP1196345/INV-008813, both funded by the Bill and Melinda Gates Foundation. This work was also supported by the National Institute for Allergy and Infectious Diseases through the Scripps Consortium for HIV Vaccine Development (CHAVD) (AI144462). A.J.B is a recipient of a Chris Scanlan Memorial Scholarship from Corpus Christi College, Oxford.
Funding Information:
We thank Dr. G. Bousfield (Wichita State University, KS) for permission to use the spectrum of Man3 GlcNAc3 GalNAc1.
Publisher Copyright:
© 2021, The Author(s).
Keywords:
Human immunodeficiency virus, Ion mobility, N-Acetylgalactosamine, N-Glycans, Negative ion fragmentation
Identifiers
Local EPrints ID: 451048
URI: http://eprints.soton.ac.uk/id/eprint/451048
ISSN: 1618-2642
PURE UUID: e2e0874e-d6a7-485b-81df-3bd8049ded85
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Date deposited: 03 Sep 2021 16:44
Last modified: 17 Mar 2024 03:47
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Contributors
Author:
David J. Harvey
Author:
Anna-Janina Behrens
Author:
Weston B. Struwe
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