purification of neutrophil collagenase and gelatinase B from CF sputum &

the modulation of collagen production in human lung fibroblasts by inflammatory cytokines
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by
University College Dublin , Dublin
Cystic fibrosis., Lungs -- Diseases., Enzymes., Collagen., Cytok
Other titlesModulation of collagen production in human lung fibroblasts by imflammatory cytokines.
StatementDawn J. Macfarlane.
ContributionsUniversity College Dublin. Department of Pharmacology.
The Physical Object
Paginationiii, 147p., [11]p. of plates :
ID Numbers
Open LibraryOL19619684M

Murphy G, Bretz U, Baggiolini M, Reynolds JJ. The latent collagenase and gelatinase of human polymorphonuclear neutrophil leucocytes. Biochem J. Nov 15; (2)– [PMC free article] Murphy G, Cawston TE, Reynolds JJ.

An inhibitor of collagenase from human amniotic fluid. Purification, characterization and action on by:   Processing of chemokines by gelatinase B and neutrophil collagenase. Gelatinase B has been found to process the CXC chemokines IL‐8, CTAP‐III, GRO‐α, PF‐4 [] and SDF‐1 [].

To complement and compare the processing of other chemokines by gelatinase B, human GCP‐2 was incubated with natural gelatinase B from human neutrophils at an Cited by: Gelatinase B/MMP-9 and neutrophil collagenase/MMP-8 process the chemokines human GCP-2/CXCL6, ENA/CXCL5 and mouse GCP-2/LIX and modulate their physiological activities Philippe E.

Van den Steen, Anja Wuyts, Steven J. Husson, Paul Proost, Jo Van Damme and Ghislain OpdenakkerCited by: secretagogue, rapid gelatinase release was observed. When granulocytes were stimulated with the neutrophil- activating peptide interleukin-8, maximal exocytosis occurred within 1 h. The almost immediate release of neutrophil gelatinase after stimulation of the cells with a chemotactic factor might play a key role in remodelingCited by: Jialiang Hu, Philippe E.

Van den Steen, Chris Dillen, Ghislain Opdenakker, Targeting neutrophil collagenase/matrix metalloproteinase-8 and gelatinase B/matrix metalloproteinase-9 with a peptidomimetic inhibitor protects against endotoxin shock, Biochemical Pharmacology, /, 70, 4, (), ().Cited by: Purification and identification of 91‐kDa neutrophil gelatinase truncated form of tumor-derived kDa gelatinase (type IV collagenase), lacking eight residues at the NH2-terminus.

Details purification of neutrophil collagenase and gelatinase B from CF sputum & FB2

Neutrophil collagenase/matrix metalloproteinase-8 (MMP-8) and gelatinase B/matrix metalloproteinase-9 (MMP-9) are contained in granules, are quickly exocytosed upon granulocyte activation and efficiently cleave intact and denatured collagens, respectively.

Genetic ablation of gelatinase B protects against endotoxin-induced mortality. a, PGP production was significantly increased in CF samples compared with normal control samples on type I collagen: CF sputum (n 10) and normal control sputum (n 10) were each incubated on. The neutrophil contains numerous granules of various composition and structure.

For decades, the neutrophil was believed to contain only two granule types, peroxisomes, or peroxidase-positive granules, and peroxidase-negative granules. Later, existence of the third type distinguished by the presence of gelatinase hydrolyzing collagen and gelatin was proposed.

Salvatore Di Somma, Rossella Marino, in Critical Care Nephrology (Third Edition), Neutrophil Gelatinase-Associated Lipocalin. Neutrophil gelatinase-associated lipocalin (NGAL) is an independent biologic marker able to detect earlier AKI than SCr.

24 In fact SCr is a marker of kidney function, whereas NGAL is a marker of kidney injury.

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25 Moreover, NGAL levels are useful to quantify the. ELSEVIER Bone Vol. 17, No. 3 September The Synthesis of Collagenase, Gelatinise-A (72 kDa) and -B (95 kDa), and TIMP-1 and -2 by Human Osteoblasts From Normal and Arthritic Bone M.

MEIKLE,1'2 S. BORD,' R. HEMBRY,1 and J. REYNOLDS' ' Cell and Molecular Biology Department, Strangeways Research Laboratory, Cambridge, UK 2 Department of. Neutrophil gelatinase B potentiates interleukin-8 tenfold by aminoter- minal processing, whereas it degrades CTAP-III, PF-4, and GRO- ␣ and leaves RANTES and MCP-2 intact.

Induced sputum IL-8 gene expression, neutrophil influx and MMP-9 in allergic bronchopulmonary aspergillosis. The collagenase activity appears to be. Human neutrophils secrete gelatinase B in. Function. Due to its role in cleaving collagen in the extracellular matrix, gelatinase B has multiple functional roles in normal physiology.

Neutrophil action. Gelatinase B, along with elastase, appears to be a regulatory factor in neutrophil migration across the basement membrane.

Gelatinase B plays several important functions within neutrophil action, such as degrading extracellular matrix. of collagenases 1 and 2, and gelatinases A and B, while collagenase 3 was not found.

Neutrophils exhibited a positive signal for collagenase 2 and gelatinase B, whereas collagenase 1 and gelatinase A were revealed mainly in macrophages and epithelial cells.

BAL gelatin zymography showed a moderate increase of progelatinase-A activity and. Investigation of collagenase and gelatinase inhibitory natural components afforded two isoflavonoids.

Two isoflavonoids, tectorigeninO-β-D-glucoside (1) and luteolinO-β-D-glucuronopyranoside (2), were isolated from ethyl acetate fraction of Viola patrinii fermentation extracts (VPFE).Of these, compounds 1 and 2 exhibited collagenase inhibitory activity (IC 50) at a concentration of. Paemen L, Martens E, Masure S, Opdenakker G.

Monoclonal antibodies specific for natural human neutrophil gelatinase B used for affinity purification, quantitation by two-site ELISA and inhibition of enzymatic activity.

Eur J Biochem. Dec 15; (3)– Collagenase is secreted from neutrophils as a latent or proenzyme. In an effort to understand the mechanism of collagenase activation in inflammation, human peripheral neutrophils (PMNs) were isolated and incubated with the tumor promotor, phorbol myristate acetate (PMA), which induces the neutrophils to degranulate and secrete proteinases.

Neutrophil media were then treated with various. Gelatinase and possibly other enzymes stored in this secretory organelle may be involved in the early events of neutrophil mobilization, the response to chemotactic signals and diapedesis.

Full text Get a printable copy (PDF file) of the complete article (M), or click on a. Patrick Henriet, Yves Eeckhout, in Handbook of Proteolytic Enzymes (Third Edition), Name and History.

Collagenase 3 is a true collagenase, i.e. a proteolytic enzyme that efficiently cleaves native triple-helical type I collagen into characteristic 3/4 and 1/4 s neutrophil collagenase (collagenase 2, MMP-8) encoded by a specific gene in human, collagenase activities in. MMP-9 [matrix metalloproteinase 9; metallopeptidase 9; MMP-9, also known as 92 kDa type IV collagenase, 92 kDa gelatinase or gelatinase B (GELB)], is expressed in a variety of cells in the lung.

Here, the mechanisms by which neutrophil collagenase (MMP-8), gelatinase A (MMP-2), and the ectodomain of MT1-MMP (ectMMP) degrade fibrillar collagen were examined.

In particular, the hydrolysis of type I collagen at 37 °C was investigated to identify functional differences in the processing of the two α–chain types of fibrillar collagen.

Neutrophil gelatinase associated lipocalin (NGAL) also known as migration stimulating factor inhibitor (MSFI), human neutrophil lipocalin (HNL), alpha-1 microglobulin related protein, siderocalin or uterocalin is a amino acid glycoprotein encoded by a.

No gelatinase activity could be found in PBMC treated with LPS during that time period (0 to 4 h). We confirmed the nature of the gelatinase identified by zymography as being MMP-9, using a Western blot technique on stimulated neutrophil supernatants (Figure 4).

We found an upregulation of a and a kD band reacting with the anti–MMP Knäuper V, López-Otin C, Smith B, Knight G, Murphy G.

Biochemical characterization of human collagenase J Biol Chem. Jan 19; (3)– Cawston TE, Barrett AJ. A rapid and reproducible assay for collagenase using [C]acetylated collagen. Anal. We found sputum from TB patients had increased MMP-8 concentrations, neutrophil myeloperoxidase (MPO) and neutrophil gelatinase associated lipocalin (NGAL) compared to controls.

MMP-8 was strongly associated with markers of neutrophil activation, MPO and NGAL, indicating that sputum MMP-8 is likely to be neutrophil-derived. Comparable to SLPI, MMP-9, also called gelatinase B, is produced and secreted by immune cells, in particular by neutrophils, and attributed antimicrobial and immune-modulatory functions.

In addition, MMP-9 is implicated in many chronic lung pathologies, including bronchiectasis (29) and cystic fibrosis.

Furthermore, because sputum trypsin activates human progelatinase B, it may be responsible in part for the reported presence of activated matrix metalloproteinases in CF sputum.

the identification and purification of an active, trypsin-like protease from purulent cystic fibrosis (CF) sputum was first reported by Viscarello and colleagues (11).

Evidence for the contribution of neutrophils to the pathogenesis of pulmonary emphysema is not convincing. We evaluated neutrophil involvement in subclinical pulmonary emphysema by measuring human neutrophil lipocalin (HNL) and two matrix metalloproteinases, gelatinase B (MMP-9) and neutrophil collagenase (MMP-8), in bronchoalveolar lavage fluid (BALF) from 65 community-based.

licheniformis, B. brevis and B. megaterium were either completely inactive or produced only partial disintegration. Collagenase production was always accom- panied by gelatinase production but the reverse was not the case. Concentrated culture filtrates of B.

cereus were shown to contain collagenase and gelatinase. antibodywasfoundto react with neutrophil collagenase in immuno-blotting or immunoprecipitation experiments. Purified IgG was ob- In previousstudies wereported the purification ofneutrophil gelatinase (7).

Description purification of neutrophil collagenase and gelatinase B from CF sputum & EPUB

Theenzymepurified to ahigh specific activity was found to .Among MMPs secreted from neutrophils may be interstitial collagenase (MMP-8) initiating the cleavage of the triple helix of native collagen types I, II, III, and X, and 92 kDa gelatinase (MMP-9) cleaving native types IV and V collagens, and to a lesser extent, fibronectin, laminin, entactin, and insoluble elastin.

Thus, with regard to collagen.To examine the clinical significance of neutrophil gelatinase in rheumatic diseases, plasma and synovial fluid (SF) gelatinase levels were determined in 62 patients with rheumatoid arthritis (RA), 12 patients with ankylosing spondylitis (AS), 18 patients with osteoarthritis (OA) and 17 healthy controls.

The gelatinase level was measured by enzyme-linked immunoassay (ELISA).