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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions |
umls-concept:C0005889,
umls-concept:C0030705,
umls-concept:C0036087,
umls-concept:C0039082,
umls-concept:C0205177,
umls-concept:C0205217,
umls-concept:C0205275,
umls-concept:C0205314,
umls-concept:C0205352,
umls-concept:C0205460,
umls-concept:C0441655,
umls-concept:C0441889,
umls-concept:C0679622,
umls-concept:C0680730,
umls-concept:C1148554,
umls-concept:C1510438,
umls-concept:C1521806,
umls-concept:C2700400
|
| pubmed:issue |
8-9
|
| pubmed:dateCreated |
1999-4-13
|
| pubmed:abstractText |
Here we describe a new principle for accessing the activity of the different members of the human matrix-metalloproteinases (MMPs) by a colorimetric assay. Using protein engineering, a modified pro-urokinase was made in which the activation sequence, normally recognized by plasmin (ProArgPheLys/IleIleGlyGly), was replaced by a sequence that is specifically recognized by MMPs (ArgProLeuGly/IleIleGlyGly). The active urokinase resulting from the activation of this modified pro-urokinase by MMPs can be measured directly using a chromogenic peptide substrate for urokinase. The assay has been made specific for MMP-9 using an MMP-9 specific monoclonal antibody. Using this antibody MMP-9 is captured from biological fluids or tissue culture media, and MMP-activity of both active and latent MMP-9 can be analysed. We determined the gelatinase-B (MMP-9) activity present in saliva from patients with Sjögren's syndrome. Using a general gelatinase assay with radioactively-labeled gelatinated collagen it was observed that gelatinase activity was slightly, though not significantly, increased in patients: general gelatinase activity in patients versus healthy controls: 17.0 +/- 4.9 vs 12.2 +/- 2.5 x 10(4) cpm/ml (p > 0.05, and 44.0 (4.0 vs 36.1 +/- 1.9 x 10(4) cpm/ml (p > 0.05), for active and latent gelatinase, respectively. However, using the immunocapture activity assay (using modified urokinase) specifically MMP-9 activity was measured, which was significantly increased in saliva from patients compared to healthy controls: MMP-9 (already active): patients 8.9 +/- 2.5 U/mg, controls 1.0 +/- 0.5 U/mg (p = 0.002); latent plus active MMP-9: patients 53.1 +/- 9.8 U/mg, controls 16.5 +/- 2.6 U/mg (p = 0.01). This assay, measuring MMP-9 activity using modified pro-urokinase as a substrate can easily be adapted for the specific detection of the various members of the MMP-family or other difficult to measure proteases, in a format that can be used for high throughput screening of compounds or samples.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Dec
|
| pubmed:issn |
0945-053X
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
17
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
657-65
|
| pubmed:dateRevised |
2006-11-15
|
| pubmed:meshHeading |
pubmed-meshheading:9923658-Animals,
pubmed-meshheading:9923658-Antibodies, Monoclonal,
pubmed-meshheading:9923658-Antibody Specificity,
pubmed-meshheading:9923658-Collagenases,
pubmed-meshheading:9923658-Female,
pubmed-meshheading:9923658-Humans,
pubmed-meshheading:9923658-Immunoassay,
pubmed-meshheading:9923658-Matrix Metalloproteinase 9,
pubmed-meshheading:9923658-Mice,
pubmed-meshheading:9923658-Mice, Inbred BALB C,
pubmed-meshheading:9923658-Saliva,
pubmed-meshheading:9923658-Sensitivity and Specificity,
pubmed-meshheading:9923658-Sjogren's Syndrome,
pubmed-meshheading:9923658-Tissue Inhibitor of Metalloproteinase-2
|
| pubmed:year |
1998
|
| pubmed:articleTitle |
A novel and simple immunocapture assay for determination of gelatinase-B (MMP-9) activities in biological fluids: saliva from patients with Sjögren's syndrome contain increased latent and active gelatinase-B levels.
|
| pubmed:affiliation |
Gaubius Laboratory TNO-PG, Leiden, The Netherlands. R.HANEMAAIJER@PG.TNO.NL
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|