Glycan analysis of human neutrophil granules implicates a maturation-dependent glycosylation machinery
Artikel i vetenskaplig tidskrift, 2020

Protein glycosylation is essential to trafficking and immune functions of human neutrophils. During granulopoiesis in the bone marrow, distinct neutrophil granules are successively formed. Distinct receptors and effector proteins, many of which are glycosylated, are targeted to each type of granule according to their time of expression, a process called "targeting by timing." Therefore, these granules are time capsules reflecting different times of maturation that can be used to understand the glycosylation process during granulopoiesis. Herein, neutrophil subcellular granules were fractionated by Percoll density gradient centrifugation, andN- andO-glycans present in each compartment were analyzed by LC-MS. We found abundant paucimannosidicN-glycans and lack ofO-glycans in the early-formed azurophil granules, whereas the later-formed specific and gelatinase granules and secretory vesicles contained complexN-andO-glycans with remarkably elongatedN-acetyllactosamine repeats with Lewis epitopes. Immunoblotting and histochemical analysis confirmed the expression of Lewis X and sialyl-Lewis X in the intracellular granules and on the cell surface, respectively. Many glycans identified are unique to neutrophils, and their complexity increased progressively from azurophil granules to specific granules and then to gelatinase granules, suggesting temporal changes in the glycosylation machinery indicative of "glycosylation by timing" during granulopoiesis. In summary, this comprehensive neutrophil granule glycome map, the first of its kind, highlights novel granule-specific glycosylation features and is a crucial first step toward a better understanding of the mechanisms regulating protein glycosylation during neutrophil granulopoiesis and a more detailed understanding of neutrophil biology and function.

mass spectrometry (MS)





Lewis epitope


plasma membrane

targeting by timing

liquid chromatography



N-linked glycosylation


Vignesh Venkatakrishnan

Göteborgs universitet

Regis Dieckmann

Göteborgs universitet

Ian Loke

Macquarie University

Cordlife Group

Harry C. Tjondro

Macquarie University

Sayantani Chatterjee

Macquarie University

Johan Bylund

Göteborgs universitet

Morten Thaysen-Andersen

Macquarie University

Niclas G. Karlsson

Göteborgs universitet

Anna Karlsson-Bengtsson

Göteborgs universitet

Chalmers, Biologi och bioteknik, Kemisk biologi

Journal of Biological Chemistry

0021-9258 (ISSN) 1083-351X (eISSN)

Vol. 295 36 12648-12660

Glykosylering i humana neutrofiler vid sepsis och systemisk inflammation

Vetenskapsrådet (VR) (2018-03077), 2019-01-01 -- 2024-12-31.


Biokemi och molekylärbiologi

Annan medicinsk grundvetenskap

Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)





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