A fluorescent screening assay for collagenase using collagen labeled with 2-methoxy-2,4-diphenyl-3(2H)-furanone

O'Grady, Robert L.; Nethery, Andrew; Hunter, Neil

doi:10.1016/0003-2697(84)90199-4

Cited by 72 publications

(28 citation statements)

References 13 publications

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“…Solution-phase assays used heat-denatured rat-tail tendon collagen (22) that had been labeled with 2-methoxy-2,4-diphenyl-3(2H)furanone (MDPF; TSI, Tokyo, Japan; Ref. 23). A total of 40 g of MDPF-labeled gelatin was incubated with the sample at 37°C overnight in MMP assay buffer.…”

Section: Analysis Of Gelatin-degrading Activitymentioning

confidence: 99%

Activation of Macrophage Promatrix Metalloproteinase-9 by Lipopolysaccharide-Associated Proteinases

Min

Moore

Bain

et al. 2002

The Journal of Immunology

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LPS induces an up-regulation of promatrix metalloproteinase-9 (proMMP9) gene expression in cells of the monocyte/macrophage lineage. We demonstrate here that LPS preparations are also able to activate proMMP9 made by human macrophages or THP-1 cells via LPS-associated proteinases, which cleave the N-terminal propeptide at a site or sites close to the one cleaved upon activation with organomercurial compounds. LPS-associated proteinases are serine proteinases that are able to cleave denatured collagens (gelatin) and the mammalian serine proteinase inhibitor, α1-proteinase inhibitor, thereby pushing the balance of extracellular matrix turnover even further toward degradation. A low molecular mass, low affinity inhibitor of MMP9, possibly derived from the propeptide, is generated during proMMP9 activation. However, inhibition of the LPS-associated proteinases had no effect on proMMP9 synthesis, indicating that their proteolytic activity was not required for signaling the up-regulation of the proMMP9 gene.

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Section: Analysis Of Gelatin-degrading Activitymentioning

confidence: 99%

Activation of Macrophage Promatrix Metalloproteinase-9 by Lipopolysaccharide-Associated Proteinases

Min

Moore

Bain

et al. 2002

The Journal of Immunology

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“…Aliquots of conditioned media were fractionated on 10% SDS ± PAGE containing 1 mg/ml gelatin (G-6269; Sigma) and 0.5 mg/ml 2-methoxy-2,4-diphenyl-3(2H)-furanone (Fluka 645989) (O'Grady et al, 1984). The gels were washed for 30 min in 50 mM Tris, 0.02% NaN 3 and 2.5% Triton X-100, pH 7.5 and for 30 min in the same bu er supplemented with 5 mM CaCl 2 and 1 mM ZnCl 2 (Heussen and Dowdle, 1980).…”

Section: Gelatinase Zymographymentioning

confidence: 99%

Inhibition of collagenase-3 (MMP-13) expression in transformed human keratinocytes by interferon-γ is associated with activation of extracellular signal-regulated kinase-1,2 and STAT1

et al. 2000

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Collagenase-3 (MMP-13) is characterized by an exceptionally wide substrate speci®city and restricted expression. MMP-13 is speci®cally expressed by transformed human keratinocytes in squamous cell carcinomas in vivo and its expression correlates with their invasion capacity. Here, we show, that interferon-g (IFN-g) markedly inhibits expression of MMP-13 by human cutaneous SCC cells (UT-SCC-7) and by ras-transformed human epidermal keratinocytes (A-5 cells) at the transcriptional level. In addition, IFN-g inhibits collagenase-1 (MMP-1) expression in these cells. IFN-g abolished the enhancement of MMP-13 and MMP-1 expression by transforming growth factor-b (TGF-b) and tumor necrosis factor-a (TNF-a), and inhibited invasion of A-5 cells through type I collagen. IFN-g also rapidly and transiently activates extracellular signal-regulated kinase 1,2 (ERK1,2) and blocking ERK1,2 pathway (Raf/MEK1,2/ERK1,2) by speci®c MEK1,2 inhibitor PD98059 partially (by 50%) prevents Ser-727 phosphorylation of STAT1 and suppression of MMP-13 expression by IFN-g. Furthermore, Ser-727 phosphorylation of STAT1 by ERK1,2, or independently of ERK1,2 activation is associated with marked reduction in MMP-13 expression. These observations identify a novel role for IFN-g as a potent inhibitor of collagenolytic activity and invasion of transformed squamous epithelial cells, and show that inhibition of MMP-13 expression by IFN-g involves activation of ERK1,2 and STAT1. Oncogene (2000) 19, 248 ± 257.

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“…(10,11)). The assay of Substrate degradation is achieved by using radioactive, fluorescent (12) or chromophore labelled Substrates, äs well äs by measuring the change in physical parameters such äs viscosity or UV-absorbance (13).…”

Section: Introductionmentioning

confidence: 99%

Enzyme Linked Immunosorbent Assays (ELISA) for the Quantitative Determination of Human Leukocyte Collagenase and Gelatinase

Bergmann¹,

Michaelis²,

Oberhoff³

et al. 1989

Clinical Chemistry and Laboratory Medicine

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Summary:A competitive and a sandwich enzyme linked immunosorbent assay (ELISA) were developed for human leukocyte collagenase and gelatinase.The competitive assay could detect 0.5 ng collagenase and 0.05 ng gelatinase. The detection limit of the sandwich ELISA was 0.05 ng for collagenase and 0.02 ng for gelatinase.No cross reactivity between human leukocyte collagenase and gelatinase was detected. The sandwich ELISA was used to determine plasma levels of these enzymes. The 90% ränge for collagenase was between 0 and 50 g/l; the 90% ränge for gelatinase was between 27 and 94 g/l.

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A fluorescent screening assay for collagenase using collagen labeled with 2-methoxy-2,4-diphenyl-3(2H)-furanone

Cited by 72 publications

References 13 publications

Activation of Macrophage Promatrix Metalloproteinase-9 by Lipopolysaccharide-Associated Proteinases

Activation of Macrophage Promatrix Metalloproteinase-9 by Lipopolysaccharide-Associated Proteinases

Inhibition of collagenase-3 (MMP-13) expression in transformed human keratinocytes by interferon-γ is associated with activation of extracellular signal-regulated kinase-1,2 and STAT1

Enzyme Linked Immunosorbent Assays (ELISA) for the Quantitative Determination of Human Leukocyte Collagenase and Gelatinase

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