N-glycan microheterogeneity regulates interactions of plasma proteins

Di Wu, Weston B Struwe, David J Harvey, Michael A J Ferguson, Carol V Robinson

Research output: Contribution to journalArticle

8 Citations (Scopus)
65 Downloads (Pure)

Abstract

Altered glycosylation patterns of plasma proteins are associated with autoimmune disorders and pathogenesis of various cancers. Elucidating glycoprotein microheterogeneity and relating subtle changes in the glycan structural repertoire to changes in protein-protein, or protein-small molecule interactions, remains a significant challenge in glycobiology. Here, we apply mass spectrometry-based approaches to elucidate the global and site-specific microheterogeneity of two plasma proteins: α1-acid glycoprotein (AGP) and haptoglobin (Hp). We then determine the dissociation constants of the anticoagulant warfarin to different AGP glycoforms and reveal how subtle N-glycan differences, namely, increased antennae branching and terminal fucosylation, reduce drug-binding affinity. Conversely, similar analysis of the haptoglobin-hemoglobin (Hp-Hb) complex reveals the contrary effects of fucosylation and N-glycan branching on Hp-Hb interactions. Taken together, our results not only elucidate how glycoprotein microheterogeneity regulates protein-drug/protein interactions but also inform the pharmacokinetics of plasma proteins, many of which are drug targets, and whose glycosylation status changes in various disease states.

Original languageEnglish
Article number 201807439
Pages (from-to)8763-8768
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume115
Issue number35
Early online date15 Aug 2018
DOIs
Publication statusPublished - 28 Aug 2018

Fingerprint

Polysaccharides
Blood Proteins
Glycoproteins
Haptoglobins
Proteins
Glycosylation
Glycomics
Acids
Warfarin
Drug Interactions
Pharmaceutical Preparations
Anticoagulants
Mass Spectrometry
Pharmacokinetics
Neoplasms

Keywords

  • glycoprotein
  • mass spectrometry
  • protein interactions
  • Glycoprotein
  • Mass spectrometry
  • Protein interactions
  • Glucans/chemistry
  • Models, Chemical
  • Humans
  • Haptoglobins/chemistry
  • Orosomucoid/chemistry
  • Warfarin/chemistry

Cite this

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title = "N-glycan microheterogeneity regulates interactions of plasma proteins",
abstract = "Altered glycosylation patterns of plasma proteins are associated with autoimmune disorders and pathogenesis of various cancers. Elucidating glycoprotein microheterogeneity and relating subtle changes in the glycan structural repertoire to changes in protein-protein, or protein-small molecule interactions, remains a significant challenge in glycobiology. Here, we apply mass spectrometry-based approaches to elucidate the global and site-specific microheterogeneity of two plasma proteins: α1-acid glycoprotein (AGP) and haptoglobin (Hp). We then determine the dissociation constants of the anticoagulant warfarin to different AGP glycoforms and reveal how subtle N-glycan differences, namely, increased antennae branching and terminal fucosylation, reduce drug-binding affinity. Conversely, similar analysis of the haptoglobin-hemoglobin (Hp-Hb) complex reveals the contrary effects of fucosylation and N-glycan branching on Hp-Hb interactions. Taken together, our results not only elucidate how glycoprotein microheterogeneity regulates protein-drug/protein interactions but also inform the pharmacokinetics of plasma proteins, many of which are drug targets, and whose glycosylation status changes in various disease states.",
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N-glycan microheterogeneity regulates interactions of plasma proteins. / Wu, Di; Struwe, Weston B; Harvey, David J; Ferguson, Michael A J; Robinson, Carol V.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 115, No. 35, 201807439, 28.08.2018, p. 8763-8768.

Research output: Contribution to journalArticle

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T1 - N-glycan microheterogeneity regulates interactions of plasma proteins

AU - Wu, Di

AU - Struwe, Weston B

AU - Harvey, David J

AU - Ferguson, Michael A J

AU - Robinson, Carol V

N1 - We thank Joseph Gault, Hsin-Yung Yen, and Manman Guo (Nuffield Department of Orthopaedics, Rheumatology, and Musculoskeletal Sciences, University of Oxford) for useful discussions in native MS, glycobiology, and glycoproteomics analysis. We acknowledge funding from Medical Research Council Programme Grant MR/N020413/1, European Research Council Advanced Grant ENABLE (641317), and Wellcome Trust Investigator Award 104633/Z/14/Z.

PY - 2018/8/28

Y1 - 2018/8/28

N2 - Altered glycosylation patterns of plasma proteins are associated with autoimmune disorders and pathogenesis of various cancers. Elucidating glycoprotein microheterogeneity and relating subtle changes in the glycan structural repertoire to changes in protein-protein, or protein-small molecule interactions, remains a significant challenge in glycobiology. Here, we apply mass spectrometry-based approaches to elucidate the global and site-specific microheterogeneity of two plasma proteins: α1-acid glycoprotein (AGP) and haptoglobin (Hp). We then determine the dissociation constants of the anticoagulant warfarin to different AGP glycoforms and reveal how subtle N-glycan differences, namely, increased antennae branching and terminal fucosylation, reduce drug-binding affinity. Conversely, similar analysis of the haptoglobin-hemoglobin (Hp-Hb) complex reveals the contrary effects of fucosylation and N-glycan branching on Hp-Hb interactions. Taken together, our results not only elucidate how glycoprotein microheterogeneity regulates protein-drug/protein interactions but also inform the pharmacokinetics of plasma proteins, many of which are drug targets, and whose glycosylation status changes in various disease states.

AB - Altered glycosylation patterns of plasma proteins are associated with autoimmune disorders and pathogenesis of various cancers. Elucidating glycoprotein microheterogeneity and relating subtle changes in the glycan structural repertoire to changes in protein-protein, or protein-small molecule interactions, remains a significant challenge in glycobiology. Here, we apply mass spectrometry-based approaches to elucidate the global and site-specific microheterogeneity of two plasma proteins: α1-acid glycoprotein (AGP) and haptoglobin (Hp). We then determine the dissociation constants of the anticoagulant warfarin to different AGP glycoforms and reveal how subtle N-glycan differences, namely, increased antennae branching and terminal fucosylation, reduce drug-binding affinity. Conversely, similar analysis of the haptoglobin-hemoglobin (Hp-Hb) complex reveals the contrary effects of fucosylation and N-glycan branching on Hp-Hb interactions. Taken together, our results not only elucidate how glycoprotein microheterogeneity regulates protein-drug/protein interactions but also inform the pharmacokinetics of plasma proteins, many of which are drug targets, and whose glycosylation status changes in various disease states.

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KW - Orosomucoid/chemistry

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