Publikation: Bidrag til tidsskriftReviewForskningfagfællebedømt

Protein glycosylation involves the co-translational or post-translational addition of glycans to proteins and is a crucial protein modification in health and disease. The aim of glycoproteomics is to understand how glycosylation shapes biological processes by understanding peptide sequences, glycan structures and sites of modification in a system-wide context. Over the past two decades, mass spectrometry (MS) has emerged as the primary technique for studying glycoproteins, with intact glycopeptide analysis — the study of glycopeptides decorated with their native glycan structures — now a preferred approach across the community. In this Primer, we discuss glycoproteomic methods for studying glycosylation classes, including best practices and critical considerations. We summarize how glycoproteomics is used to understand glycosylation at a systems level, with a specific focus on N-linked and O-linked glycosylation (both mucin-type and O-GlcNAcylation). We cover topics that include sample selection; techniques for protein isolation, proteolytic digestion, glycopeptide enrichment and MS fragmentation; bioinformatic platforms and applications of glycoproteomics. Finally, we give a perspective on where the field is heading. Overall, this Primer outlines the current technologies, persistent challenges and recent advances in the exciting field of glycoproteomics.

TidsskriftNature Reviews Methods Primers
Udgave nummer1
StatusUdgivet - 2022

Bibliografisk note

Funding Information:
N.E.S. is supported by an Australian Research Council Future Fellowship (FT200100270). B.L.P. is supported by an Australian National Health and Medical Research Emerging Leader Grant (APP 2009642). M.T.-A. is supported by an Australian Research Council Future Fellowship (FT210100455). D.A.P. and A.I.N. are supported by NIH grants R01-GM-094231 and U24-CA210967. S.A.M. is supported by the Yale Science Development Fund and the Yale SEAS/Science Program to Advance Research Collaboration (SPARC). A.H. is supported by a research grant (00025438) from VILLUM FONDEN. K.S. is supported by an Excellence grant from the Novo Nordisk Foundation (NNF17OC0026030). H.H.W. is supported by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (GlycoSkin H2020-ERC; 772735), Lundbeck Foundation (R313-2019-869) and the Neye Foundation. The Copenhagen Center for Glycomics including I.B., K.S., A.H., H.H.W. and S.Y.V. are supported by a Danish National Research Foundation grant (DNRF107). N.M.R. acknowledges support from a NIH Predoctoral to Postdoctoral Transition award (K00 CA212454). K.F.A.-K. is supported by the Japan Science and Technology Agency (JST) and National Bioscience Database Center (NBDC) of Japan, grant number 17934031.

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