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Hyper-truncated Asn355- and Asn391-glycans modulate the activity of neutrophil granule myeloperoxidase.
Tjondro, Harry C; Ugonotti, Julian; Kawahara, Rebeca; Chatterjee, Sayantani; Loke, Ian; Chen, Siyun; Soltermann, Fabian; Hinneburg, Hannes; Parker, Benjamin L; Venkatakrishnan, Vignesh; Dieckmann, Regis; Grant, Oliver C; Bylund, Johan; Rodger, Alison; Woods, Robert J; Karlsson-Bengtsson, Anna; Struwe, Weston B; Thaysen-Andersen, Morten.
Affiliation
  • Tjondro HC; Department of Molecular Sciences, Macquarie University, Sydney, New South Wales, Australia; Biomolecular Discovery Research Centre, Macquarie University, Sydney, New South Wales, Australia.
  • Ugonotti J; Department of Molecular Sciences, Macquarie University, Sydney, New South Wales, Australia; Biomolecular Discovery Research Centre, Macquarie University, Sydney, New South Wales, Australia.
  • Kawahara R; Department of Molecular Sciences, Macquarie University, Sydney, New South Wales, Australia; Biomolecular Discovery Research Centre, Macquarie University, Sydney, New South Wales, Australia.
  • Chatterjee S; Department of Molecular Sciences, Macquarie University, Sydney, New South Wales, Australia; Biomolecular Discovery Research Centre, Macquarie University, Sydney, New South Wales, Australia.
  • Loke I; Cordlife Group Limited, Singapore, Singapore.
  • Chen S; Department of Chemistry, University of Oxford, Oxford, United Kingdom.
  • Soltermann F; Department of Chemistry, University of Oxford, Oxford, United Kingdom.
  • Hinneburg H; Department of Molecular Sciences, Macquarie University, Sydney, New South Wales, Australia; Biomolecular Discovery Research Centre, Macquarie University, Sydney, New South Wales, Australia.
  • Parker BL; Department of Physiology, University of Melbourne, Melbourne, Victoria, Australia.
  • Venkatakrishnan V; Department of Rheumatology and Inflammation Research, Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
  • Dieckmann R; Department of Rheumatology and Inflammation Research, Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
  • Grant OC; Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia, USA.
  • Bylund J; Department of Oral Microbiology and Immunology, Institute of Odontology, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
  • Rodger A; Department of Molecular Sciences, Macquarie University, Sydney, New South Wales, Australia; Biomolecular Discovery Research Centre, Macquarie University, Sydney, New South Wales, Australia.
  • Woods RJ; Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia, USA.
  • Karlsson-Bengtsson A; Department of Rheumatology and Inflammation Research, Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden; Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden.
  • Struwe WB; Department of Chemistry, University of Oxford, Oxford, United Kingdom.
  • Thaysen-Andersen M; Department of Molecular Sciences, Macquarie University, Sydney, New South Wales, Australia; Biomolecular Discovery Research Centre, Macquarie University, Sydney, New South Wales, Australia. Electronic address: morten.andersen@mq.edu.au.
J Biol Chem ; 296: 100144, 2021.
Article in En | MEDLINE | ID: mdl-33273015
ABSTRACT
Myeloperoxidase (MPO) plays essential roles in neutrophil-mediated immunity via the generation of reactive oxidation products. Complex carbohydrates decorate MPO at discrete sites, but their functional relevance remains elusive. To this end, we have characterised the structure-biosynthesis-activity relationship of neutrophil MPO (nMPO). Mass spectrometry demonstrated that nMPO carries both characteristic under-processed and hyper-truncated glycans. Occlusion of the Asn355/Asn391-glycosylation sites and the Asn323-/Asn483-glycans, located in the MPO dimerisation zone, was found to affect the local glycan processing, thereby providing a molecular basis of the site-specific nMPO glycosylation. Native mass spectrometry, mass photometry and glycopeptide profiling revealed significant molecular complexity of diprotomeric nMPO arising from heterogeneous glycosylation, oxidation, chlorination and polypeptide truncation variants and a previously unreported low-abundance monoprotomer. Longitudinal profiling of maturing, mature, granule-separated and pathogen-stimulated neutrophils demonstrated that nMPO is dynamically expressed during granulopoiesis, unevenly distributed across granules and degranulated upon activation. We also show that proMPO-to-MPO maturation occurs during early/mid-stage granulopoiesis. While similar global MPO glycosylation was observed across conditions, the conserved Asn355-/Asn391-sites displayed elevated glycan hyper-truncation, which correlated with higher enzyme activities of MPO in distinct granule populations. Enzymatic trimming of the Asn355-/Asn391-glycans recapitulated the activity gain and showed that nMPO carrying hyper-truncated glycans at these positions exhibits increased thermal stability, polypeptide accessibility and ceruloplasmin-mediated inhibition potential relative to native nMPO. Finally, molecular modelling revealed that hyper-truncated Asn355-glycans positioned in the MPO-ceruloplasmin interface are critical for uninterrupted inhibition. Here, through an innovative and comprehensive approach, we report novel functional roles of MPO glycans, providing new insight into neutrophil-mediated immunity.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Polysaccharides / Glycopeptides / Peroxidase / Cytoplasmic Granules / Neutrophils Type of study: Prognostic_studies Limits: Humans Language: En Journal: J Biol Chem Year: 2021 Document type: Article Affiliation country: Australia
Fulltext
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Polysaccharides / Glycopeptides / Peroxidase / Cytoplasmic Granules / Neutrophils Type of study: Prognostic_studies Limits: Humans Language: En Journal: J Biol Chem Year: 2021 Document type: Article Affiliation country: Australia