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2015 ; 290
(14
): 8789-802
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Human neutrophils secrete bioactive paucimannosidic proteins from azurophilic
granules into pathogen-infected sputum
#MMPMID25645918
Thaysen-Andersen M
; Venkatakrishnan V
; Loke I
; Laurini C
; Diestel S
; Parker BL
; Packer NH
J Biol Chem
2015[Apr]; 290
(14
): 8789-802
PMID25645918
show ga
Unlike plants and invertebrates, mammals reportedly lack proteins displaying
asparagine (N)-linked paucimannosylation
(mannose(1-3)fucose(0-1)N-acetylglucosamine(2)Asn). Enabled by technology
advancements in system-wide biomolecular characterization, we document that
protein paucimannosylation is a significant host-derived molecular signature of
neutrophil-rich sputum from pathogen-infected human lungs and is negligible in
pathogen-free sputum. Five types of paucimannosidic N-glycans were carried by
compartment-specific and inflammation-associated proteins of the azurophilic
granules of human neutrophils including myeloperoxidase (MPO), azurocidin, and
neutrophil elastase. The timely expressed human azurophilic granule-resident
?-hexosaminidase A displayed the capacity to generate paucimannosidic N-glycans
by trimming hybrid/complex type N-glycan intermediates with relative broad
substrate specificity. Paucimannosidic N-glycoepitopes showed significant
co-localization with ?-hexosaminidase A and the azurophilic marker MPO in human
neutrophils using immunocytochemistry. Furthermore, promyelocyte stage-specific
expression of genes coding for paucimannosidic proteins and biosynthetic enzymes
indicated a novel spatio-temporal biosynthetic route in early neutrophil
maturation. The absence of bacterial exoglycosidase activities and
paucimannosidic N-glycans excluded exogenous origins of paucimannosylation.
Paucimannosidic proteins from isolated and sputum neutrophils were preferentially
secreted upon inoculation with virulent Pseudomonas aeruginosa. Finally,
paucimannosidic proteins displayed affinities to mannose-binding lectin,
suggesting immune-related functions of paucimannosylation in activated human
neutrophils. In conclusion, we are the first to document that human neutrophils
produce, store and, upon activation, selectively secrete bioactive
paucimannosidic proteins into sputum of lungs undergoing pathogen-based
inflammation.