PXD021131

PXD021131 is an original dataset announced via ProteomeXchange.

Title	The Bimodal Sugar Code of Neutrophil Myeloperoxidase
Description	Myeloperoxidase (MPO), an important diprotomeric glycoprotein in neutrophil-mediated immunity, produces microbicidal hypohalous acids, but the underpinning glycobiology remains elusive. Deep characterisation of neutrophil-derived MPO (nMPO) using advanced mass spectrometry demonstrated that under-processed oligomannosidic- and hyper-truncated paucimannosidic- and N-acetyl-β-D-glucosamine (GlcNAc) core-type asparagine-linked glycans decorate the protein. Occlusion of Asn355 and Asn391 and sterical hindrance of Asn323- and Asn483-glycans located in the MPO dimerisation zone were found to shape the local glycan processing thereby providing a molecular basis of the site-specific nMPO glycosylation. Native mass spectrometry, mass photometry, and glycopeptide profiling revealed extreme molecular complexity of diprotomeric nMPO arising from heterogeneous glycosylation, oxidation, chlorination and polypeptide truncation variants, and a lower-abundance monomer. Longitudinal profiling of maturing, mature, granule-separated, and pathogen-activated neutrophils demonstrated that MPO is dynamically expressed during granulopoiesis, unevenly distributed across granules and rapidly degranulated, but surprisingly carries uniform glycosylation across conditions. Complete proMPO-to-MPO maturation evidently occur during early/mid-stage granulopoiesis. The conserved Asn355- and Asn391-sequons displayed elevated GlcNAc signatures and higher oxidation and chlorination activity of the secretory vesicle/plasma membrane-resident MPO relative to MPO from other granules. Endoglycosidase H-treated nMPO displaying Asn355-/Asn391-GlcNAcylation recapitulated the activity gain and showed increased thermal stability and polypeptide accessibility relative to untreated nMPO as measured by activity assays, circular dichroism and molecular dynamics. Endoglycosidase H-treated nMPO also demonstrated elevated ceruloplasmin-mediated inhibition relative to nMPO. Modelling revealed that hyper-truncated Asn355-glycans positioned in the MPO:ceruloplasmin interface are critical for uninterrupted inhibition. We report on novel bimodal roles of the peculiar MPO glycosylation providing new insight into neutrophil glycobiology.
HostingRepository	PRIDE
AnnounceDate	2021-03-07
AnnouncementXML	Submission_2021-03-07_14:38:00.484.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Rebeca Sakuma
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	dihydroxylated residue; monohydroxylated residue; N-linked glycan core; iodoacetamide derivatized residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2020-08-26 02:21:38	ID requested
⏵ 1	2021-03-07 14:38:01	announced

Tjondro HC, Ugonotti J, Kawahara R, Chatterjee S, Loke I, Chen S, Soltermann F, Hinneburg H, Parker BL, Venkatakrishnan V, Dieckmann R, Grant OC, Bylund J, Rodger A, Woods RJ, Karlsson-Bengtsson A, Struwe WB, Thaysen-Andersen M, Hyper-truncated Asn355- and Asn391-glycans modulate the activity of neutrophil granule myeloperoxidase. J Biol Chem, 296():100144(2021) [pubmed]

submitter keyword: : Myeloperoxidase, N-glycosylation, N-acetyl-β-D-glucosamine, neutrophil, granule, biosynthesis, activity, inhibition, ceruloplasmin, granulopoiesis, degranulation

Morten Thaysen-Andersen
contact affiliation	Department of Molecular Sciences, Macquarie University, Sydney, NSW, Australia
contact email	morten.andersen@mq.edu.au
lab head
Rebeca Sakuma
contact affiliation	University of Sao Paulo
contact email	rebecasakuma@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD021131
     1. Label: PRIDE project
     2. Name: The Bimodal Sugar Code of Neutrophil Myeloperoxidase