Molecular cloning of human synovial cell collagenase and selection of a single gene from genomic DNA.

Abstract

We used a subclone of a rabbit genomic clone for collagenase that cross-hybridizes with human synovial cell messenger RNA (mRNA) to identify a human collagenase complementary DNA (cDNA) clone. The human cDNA clone is 2.1 kilobases (kb) and selects a mRNA transcript of approximately the same size from primary cultures of rheumatoid synovial cells that produce collagenase, but no mRNA is selected from control (nonproducing) synovial fibroblasts. Restriction enzyme analysis and DNA sequence data indicate that our cDNA clone is full length and that it is identical to that recently described for human skin fibroblast collagenase. The cDNA clone identified a single collagenase gene of 17 kb from blots of human genomic DNA. The identity of human skin and synovial cell collagenase and the ubiquity of this enzyme and of its substrates, the interstitial collagens types I, II, and III, imply that common mechanisms controlling collagenolysis throughout the human body may be operative in both normal and disease states.

Urine proteomic analysis differentiates cholangiocarcinoma from primary sclerosing cholangitis and other benign biliary disorders

“…We performed immunostaining of liver histology sections from CC and PSC patients to confirm our hypothesis that collagen type I fragments derive from the liver through enhanced MMP activity, especially by MMP-1 20 21. We analysed differential MMP-1 expression in liver biopsy sections of six CC and six PSC patients.…”

“…The high abundance of specific collagen fragments in urine of CC patients together with the detection of several of these peptides also in blood led us to analyse expression of MMP-1 in liver sections of CC patients at the sites of tumour. We focused on MMP-1 since this is the only protease able to initiate breakdown of interstitial collagens 20 21. We observed increased expression of MMP-1 mainly on the surface of epithelial cells at the CC tumour sites compared with PSC controls.…”

Altered production of extra-cellular matrix components by muscle-derived Duchenne muscular dystrophy fibroblasts before and after TGF-β1 treatment

“…MMP-1 plays an important role in limiting fibrosis by degrading type I and III collagen fibrils (Brinckerhoff et al 1987). In the early stages of fibrosis, a slight increase in collagen III has been reported, whereas collagen I is highly increased and remains the major collagen type later on (Eckes et al 2000).…”

Transcriptional repression of the human collagenase-1 (MMP-1) gene in MDA231 breast cancer cells by all-trans-retinoic acid requires distal regions of the promoter

“…This digestion releases a 4.6-kb genomic fragment which has the same 5′ end as the full-length promoter construct. A 120-bp EcoRI/XbaI fragment corresponding to the N-terminal region of the collagenase cDNA (Brinckerhoff et al, 1987) was used as a probe for Southern blot assays. Culture medium (1 ml) was subjected to TCA precipitation and Western analysis with sheep antiserum specific to human collagenase-1.…”