title: Proteinase 3 carries small unusual carbohydrates and associates with αlpha-defensins.
abstract: The neutrophil granulocyte is an important first line of defense against intruding pathogens and it contains a range of granules armed with antibacterial peptides and proteins. Proteinase 3 (PR3) is one among several serine proteases of the azurophilic granules in neutrophil granulocytes. Here, we characterize the glycosylation of PR3 and its association with antimicrobial human neutrophil peptides (HNPs, α-defensins) and the effect of these on the mechanism of inhibition of the major plasma inhibitor of PR3, α1-antitrypsin. The glycosylation of purified, mature PR3 showed some heterogeneity with carbohydrates at Asn 102 and 147 carrying unusual small moieties indicating heavy processing. Mass spectrometric analysis and immuno blotting revealed strong association of highly purified PR3 with α-defensins and oligomers hereof. Irreversible inhibition of PR3 by α1-antitrypsin did not affect its association with defensins. Other proteins from neutrophil granules were also found to be associated with defensins, whereas purified plasma proteins did not carry defensins. These results point to a role of defensins in controlling and targeting the activity of neutrophil granule proteins.
authors.affiliation: Department of Clinical Biochemistry and Immunology, Statens Serum Institut, DK-2300 Copenhagen S., Denmark.
authors.forename: Morten
authors.initials: M
authors.lastname: Zoega
authors.forename: Tina
authors.initials: T
authors.lastname: Ravnsborg
authors.forename: Peter
authors.initials: P
authors.lastname: Højrup
authors.forename: Gunnar
authors.initials: G
authors.lastname: Houen
authors.forename: Christian
authors.initials: C
authors.lastname: Schou
mesh_terms: D050260:Carbohydrate Metabolism / Q000502:physiology; D002241:Carbohydrates / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D013058:Mass Spectrometry; D053146:Myeloblastin / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D009504:Neutrophils / Q000737:chemistry / Q000201:enzymology; D023084:alpha-Defensins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D002241:Carbohydrates; D023084:alpha-Defensins; D053146:Myeloblastin