Characterization of the biosynthesis, processing, and sorting of human HBP/CAP37/azurocidin

Lindmark, Anders; Garwicz, Daniel; Rasmussen, P. B.; Flodgaard, Hans; Gullberg, Urban

doi:10.1002/jlb.66.4.634

Cited by 41 publications

(38 citation statements)

References 50 publications

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“…40 Unfortunately, HBP was not included in this study. However, the finding by Lindmark et al 24 that HBP is not expressed at high levels in the promyelocytic HL-60 and NB4 cells might indicate that the protein is also expressed at a later stage of neutrophil development. This observation is in line with our results showing that HL-60 cells contain lower amounts of HBP than mature neutrophils.…”

Section: Discussionmentioning

confidence: 90%

“…24,[37][38][39][40] It is thought that a granule contains proteins synthesized when a compartment is formed. 5,37,39 According to this concept, azurophilic granules are already present in myeloblasts and promyelocytes.…”

Section: Discussionmentioning

confidence: 99%

“…Mouse monoclonal antibody 2F23C3 and rabbit antiserum (409A) to recombinant HBP were prepared and purified as described earlier. 24 Sheep antibody against human elastase was from The Binding Site (Birmingham, England), and rabbit antibody against human cathepsin G was from Athens Research and Technology (Athens, GA). Monoclonal antibodies against human elastase and proteinase 3 were from Research Diagnostics (Flanders, NJ) and Wieslab AB (Lund, Sweden), respectively.…”

Section: Methodsmentioning

confidence: 99%

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Secretion of heparin-binding protein from human neutrophils is determined by its localization in azurophilic granules and secretory vesicles

Tapper

Karlsson

Mörgelin

et al. 2002

Blood

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156

172

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Human neutrophils have an important role in host defense against microbial infection. At different stages of an infectious process, neutrophils progressively up-regulate receptors and release various effector molecules. These are stored in several distinct types of granules with varying propensity to be secreted. Heparin-binding protein (HBP), also known as CAP37 or azurocidin, is a multifunctional, inactive serine-protease homologue. The present work shows that HBP is released from neutrophils on stimulation with secretagogues that do not trigger the secretion of azurophilic granule content. Therefore, the subcellular localization of HBP was investigated in more detail. Immunofluorescence microscopy revealed that HBP was localized close to the plasma membrane. Further analysis by fractionation of postnuclear supernatants from cavitated neutrophils showed that HBP is stored in azurophilic granules and secretory vesicles but that it is also detected to a minor extent in the plasma membrane. IntroductionPolymorphonuclear leukocytes (PMNs) have an important role in early host defense against invading microorganisms (for reviews, see references 1 and 2). Recruitment of these cells from the bloodstream to a site of infection involves their recognition of inflammatory mediators, their binding to adhesion molecules of the vascular endothelium, and their migration across the endothelial barrier. 3 How efficiently neutrophils perform these tasks depends on a sophisticated mobilization mechanism that triggers the release of granule contents and the concomitant up-regulation of various receptors to the plasma membrane. 4 Secretory processes are also important for the extravascular migration of neutrophils through tissues. Once the cells have reached the focus of infection, they are fully activated and are able to fight the infection by secreting reactive oxygen intermediates, antimicrobial peptides, and degradative enzymes. 2 These substances can be preferentially targeted to phagosome compartments to achieve efficient killing and degradation of internalized microorganisms.Lately, much interest has been focused on the various granule types of neutrophils and their sequential mobilization during the inflammatory process (for review, see reference 5). Analysis of these granules by electron microscopy and subcellular fractionation has demonstrated that neutrophils have at least 4 different granule or vesicle types. [6][7][8][9] These are the primary or azurophilic granules that contain myeloperoxidase (MPO), bactericidal proteins, and proteinases; the secondary or specific granules that store lactoferrin and enzymes such as collagenase and gelatinase; the tertiary or gelatinase granules that, like specific granules, contain tissuedegrading enzymes; and the secretory vesicles, an easily mobilizable compartment, that contain alkaline phosphatase and plasma proteins such as human serum albumin. The 4 granule types are mobilized at different stages of the inflammatory process; secretory vesicles are more readily secreted than th...

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Section: Discussionmentioning

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Secretion of heparin-binding protein from human neutrophils is determined by its localization in azurophilic granules and secretory vesicles

Tapper

Karlsson

Mörgelin

et al. 2002

Blood

Self Cite

156

172

View full text Add to dashboard Cite

show abstract

“…Our hypothesis was that in the absence of cleavage of the N-terminal 2 amino acids by CTSC, the serine proteases would escape sorting to granules and instead be secreted constitutively to the medium. This hypothesis was founded on previous publications demonstrating that unprocessed serine proteases are routed to medium during pulsechase examination of synthesis of serine proteases in myeloid cell lines (10,(35)(36)(37). To test this hypothesis, myeloid cells were isolated from the patient's bone marrow and separated on Lymphoprep.…”

Section: Resultsmentioning

confidence: 99%

Papillon-Lefèvre syndrome patient reveals species-dependent requirements for neutrophil defenses

Sørensen¹,

Clemmensen²,

Dahl³

et al. 2014

J. Clin. Invest.

146

173

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Papillon-Lefèvre syndrome (PLS) results from mutations that inactivate cysteine protease cathepsin C (CTSC), which processes a variety of serine proteases considered essential for antimicrobial defense. Despite serine protease-deficient immune cell populations, PLS patients do not exhibit marked immunodeficiency. Here, we characterized a 24-year-old woman who had suffered from severe juvenile periodontal disease, but was otherwise healthy, and identified a homozygous missense mutation in CTSC indicative of PLS. Proteome analysis of patient neutrophil granules revealed that several proteins that normally localize to azurophil granules, including the major serine proteases, elastase, cathepsin G, and proteinase 3, were absent. Accordingly, neutrophils from this patient were incapable of producing neutrophil extracellular traps (NETs) in response to ROS and were unable to process endogenous cathelicidin hCAP-18 into the antibacterial peptide LL-37 in response to ionomycin. In immature myeloid cells from patient bone marrow, biosynthesis of CTSC and neutrophil serine proteases appeared normal along with initial processing and sorting to cellular storage. In contrast, these proteins were completely absent in mature neutrophils, indicating that CTSC mutation promotes protease degradation in more mature hematopoietic subsets, but does not affect protease production in progenitor cells. Together, these data indicate CTSC protects serine proteases from degradation in mature immune cells and suggest that neutrophil serine proteases are dispensable for human immunoprotection.

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“…They belong to the hematopoietic serine protease superfamily which also include granzymes A, B, H, K, and M in cytotoxic T-cells/NK-cells (3), and mast cell chymase (4). They all have a high degree of sequence homology, and share the same principal processing, but the individual proteases also seem to be subjected to subtle differences in this processing (1,(5)(6)(7). They are synthesized as pre-proenzymes with a signal sequence which is cleaved off immediately after translation/insertion into ER, and an N-terminal dipropeptide, which is cleaved off during processing to give an active enzyme stored in the azurophil granules (1,5).…”

Section: Introductionmentioning

confidence: 99%

The N-terminal tetrapeptide of neutrophil proteinase 3 causes S-phase arrest in granulopoietic progenitors

Sköld

Rosberg

Olofsson

2005

Experimental Hematology

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Objective: Secreted enzymatically inactive proforms of hematopoietic serine proteases proteinase 3 (PR3), azurocidin, and granzymes A, B, H, K, and M, are able to reduce the fraction of granulopoietic progenitors (CFU-GM) in S-phase, whereas human leukocyte elastase (HLE) and cathepsin G lack this ability. The objective of the present study was to map the specific sequence(s) of PR3 and other hematopoietic serine proteases responsible for the downmodulation of S-phase. Conclusion: These findings demonstrate that the downmodulatory effect on CFU-GM in S-phase is an Sphase arrest mediated by the first four N-terminal amino acids of PR3, and also suggest that this activity is dependent on the configuration of the proform providing the correct presentation of this N-terminal motif.

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Characterization of the biosynthesis, processing, and sorting of human HBP/CAP37/azurocidin

Cited by 41 publications

References 50 publications

Secretion of heparin-binding protein from human neutrophils is determined by its localization in azurophilic granules and secretory vesicles

Secretion of heparin-binding protein from human neutrophils is determined by its localization in azurophilic granules and secretory vesicles

Papillon-Lefèvre syndrome patient reveals species-dependent requirements for neutrophil defenses

The N-terminal tetrapeptide of neutrophil proteinase 3 causes S-phase arrest in granulopoietic progenitors

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