Proteinase 3 carries small unusual carbohydrates and associates with αlpha-defensins

Morten Zoega, Tina Ravnsborg, Peter Højrup, Gunnar Houen, Christian Schou

Research output: Contribution to journalJournal articleResearchpeer-review

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.
Original languageEnglish
JournalJournal of Proteomics
Volume75
Issue number5
Pages (from-to)1472-1485
ISSN1874-3919
DOIs
Publication statusPublished - 2012

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Myeloblastin
Defensins
Carbohydrates
Neutrophils
Glycosylation
Association reactions
Peptides
Proteins
Serine Proteases
Pathogens
Oligomers
Blood Proteins
Plasmas
Processing

Cite this

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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.",
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Proteinase 3 carries small unusual carbohydrates and associates with αlpha-defensins. / Zoega, Morten; Ravnsborg, Tina; Højrup, Peter; Houen, Gunnar; Schou, Christian.

In: Journal of Proteomics, Vol. 75, No. 5, 2012, p. 1472-1485.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Proteinase 3 carries small unusual carbohydrates and associates with αlpha-defensins

AU - Zoega, Morten

AU - Ravnsborg, Tina

AU - Højrup, Peter

AU - Houen, Gunnar

AU - Schou, Christian

N1 - Copyright © 2011 Elsevier B.V. All rights reserved.

PY - 2012

Y1 - 2012

N2 - 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.

AB - 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.

U2 - 10.1016/j.jprot.2011.11.019

DO - 10.1016/j.jprot.2011.11.019

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VL - 75

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JO - Journal of Proteomics

JF - Journal of Proteomics

SN - 1874-3919

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