Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.24.3 (collagenase)
 18,340 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Normal human breast organoids, derived by collagenase digestion of reduction mammaplasty tissue specimens, have been cultured in vitro for up to 28 days after injection into organ cultures of virgin rat mammary fat pads. The culture medium was serum-free Waymouth's MB 752/1 with hormonal additives. The rat mammary tissue responded well to growth-promoting and lactogenic stimuli in the culture medium, in agreement with previous investigations. Using immunohistochemistry casein was identified in rat epithelia exposed to lactogenic medium. Human organoids in culture remained viable but did not show hormone-responsiveness. Electron microscopy confirmed the presence of both luminal epithelial cells and myoepithelial cells. The serum-free culture of normal human breast organoids in a three-dimensional matrix provides a system in which to study factors controlling growth and differentiation.
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PMID:Maintenance of normal human breast organoids within rat mammary fat pads in organ culture. 310 68

Protein synthesis in the rat mammary gland has been studied using acini isolated from mammary tissue by collagenase digestion. When the acini were incubated with radioactively labeled amino acids, both cellular and milk proteins were synthesized and milk proteins were secreted into the incubation medium. Antisera to the lipogenic enzyme, fatty acid synthase, and the milk proteins, alpha-lactalbumin and the caseins, raised in rabbits, were shown to be specific by analyzing immunoprecipitates on sodium dodecyl sulfate--polyacrylamide gels. The rates of synthesis and secretion of each protein by acini prepared from rats during late gestation and at specific stages of lactation reflect their previously observed concentration in the mammary gland or milk of rats at the corresponding stage of gestation or lactation. Rats were treated according to one of the following regimes between d 7 and 14 of lactation: they were fed a control (20% casein) or a low protein (10% casein) diet ad libitum, they were fed the control diet restricted to 25 g/d (40% of the voluntary intake), they were fed the control diet for 5 d and starved for 48 h or they were treated as in 3 and then refed the control diet ad libitum for 24 h. Food restriction and starvation both resulted in lowered rates of synthesis of all proteins examined compared with either the control or refed animals. Starvation also lowered the rates of secretion of the milk proteins. Consumption of the low protein diet caused a specific decrease in both the rates of synthesis and secretion of alpha-lactalbumin compared with the control rats without affecting the synthesis and secretion of the caseins.
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PMID:Protein synthesis in mammary acini isolated from lactating rats: effect of maternal diet. 358 28

Articular chondrocytes and synovial cells were stimulated to produce collagenase, neutral casein and proteoglycan-degrading proteinases by conditioned medium from human peripheral blood mononuclear cells. Collagenase, neutral casein and proteoglycan-degrading proteinase secretion was inhibited by SR 41319, a new bisphosphonate, in a concentration-dependent manner. Complete inhibition was achieved at about 0.3 mM. EHDP exhibited the same general profile but was about 10-fold less active and never completely inhibited the enzyme secretion. When added before MCF, SR 41319 had a protective effect against subsequent activation of the cells by MCF. SR 41319 also inhibited the increase of enzyme secretion by cells previously stimulated with MCF. The results suggest that the ability of SR 41319 to inhibit the MCF-mediated secretion of neutral enzymes involved in cartilage destruction could be valuable in the management of connective tissue damage in rheumatoid arthritis.
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PMID:Effects of 1-hydroxyethylidene-1,1 bisphosphonate and (chloro-4 phenyl) thiomethylene bisphosphonic acid (SR 41319) on the mononuclear cell factor-mediated release of neutral proteinases by articular chondrocytes and synovial cells. 393 18

Stimulation of synovial cell prostaglandin production by a factor obtained from casein-induced peritoneal polymorphonuclear (PMN) cells has been investigated. Both the extract and short time cultured medium of rat peritoneal PMN cells stimulate prostaglandin (PG)E2 production as well as collagenase production in the culture of rat synovial cells. PGE2 production by the cells in the presence of the PMN factor is much faster (5 to 24 hr) than collagenase production (24 hr or later, Biomedical Res. 3, 506-516, 1982). This stimulating factor is confirmed to be derived from PMN cells, based on the purification of the cells from peritoneal exudate cells by the Ficoll-Urographin method. Elution profile of the factor on gel filtration has indicated that both PGE2 and collagenase productions by synovial cells are stimulated by the same effluent fractions corresponding to molecular weights of 15,000 - 20,000 daltons and 30,000 - 40,000 daltons. These results suggest that PMN cells are involved in PG production as well as collagenase production in the inflamed tissue by stimulating connective tissue cells such as synovial cells.
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PMID:Stimulation of prostaglandin synthesis in cultured rat synovial cells by a factor derived from polymorphonuclear leukocytes. 608 18

Six collagenases present in the culture filtrate of Clostridium histolyticum have been purified to homogeneity. Chromatography over hydroxylapatite, Sephacryl S-200, and L-arginine-Affi-Gel 202 removes the brown pigment and the great majority of the contaminating proteinases active against casein, benzoyl-L-arginine ethyl ester, and elastin. Reactive Red 120 dye ligand chromatography subdivides the collagenases, which have very similar physicochemical properties, among four fractions. The final purification is achieved by chromatography over DEAE-cellulose and SP-Sephadex. All six collagenases, designated alpha, beta, gamma, delta, epsilon and zeta by the order of their purification, are highly active against collagen and devoid of other proteolytic activities. Each exhibits a single band on sodium dodecyl sulfate-polyacrylamide gels. Two distinct subspecies of the alpha and gamma enzymes have been isolated, which have the same molecular weight and activity but different isoelectric points. There is some less pronounced microheterogeneity for the other collagenases. On the basis of their activities toward native collagen and the synthetic peptide 2-furanacryloyl-L-leucylglycyl-L-prolyl-L-alanine (FALGPA), the six collagenases are divided into two classes. Class I collagenases (alpha, beta, and gamma) have high collagenase activity and moderate FALGPA activity while the class II collagenases (sigma, epsilon, and sigma) have moderate collagenase and high FALGPA activities. The relationship between these six collagenases and other reported to have been isolated in the literature has also been examined.
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PMID:Purification and separation of individual collagenases of Clostridium histolyticum using red dye ligand chromatography. 608 87

The sensitivity and utility of proteinase assays employing fluorescamine, a compound which reacts with primary amines to form a fluorescent adduct, was assessed. As little as 1 ng of purified trypsin or clostridiopeptidase A could be detected within 3 h of incubation at 37 degrees C, using casein or gelatin as substrates. Increasing the incubation period to 18 h permitted the detection of 250 pg of each enzyme. When gelled collagen was utilized as substrate, the sensitivity to clostridiopeptidase A was reduced to 2.5 ng at 3 h and 500 pg at 18 h. The techniques could be used to measure the gelatinase, caseinase, and collagenase activities of culture media conditioned by synovial tissue. The main disadvantage of this assay is its susceptibility to interference by compounds which fluoresce or quench. This, in turn, necessitates additional blanks, which may render the assay tedious.
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PMID:An evaluation of fluorometric proteinase assays which employ fluorescamine. 609 62

A neutral protease with a marked specificity for gelatin as a protein substrate has been purified to homogeneity from medium of human skin in serum-free explant culture. The pH optimum of this gelatinase is between 7.0 and 7.5 with little or no activity displayed below pH 5. Inhibition by EDTA, ethylene glycol bis(beta-amino-ethyl ether)N,N,N',N'-tetraacetic acid (EGTA), and 1,10-phenanthroline suggest that the enzyme is a metalloendopeptidase. Calcium concentration-dependent inhibition of the enzyme by EGTA and EDTA suggest further that there is a requirement for extrinsic calcium. Indeed, removal of calcium and reduces enzyme activity, and subsequent addition of calcium restores full activity. The gelatinase is not inhibited by serine protease inhibitors but is inhibited by cysteine, dithiothreitol, and beta-mercaptoethanol. It is also inhibited by a macromolecular inhibitor of collagenase which has been purified from human skin fibroblasts. The apparent molecular weight of this enzyme, as determined by gel filtration is 120,000-150,000. The enzyme is a glycoprotein, as indicated by staining with periodic acid-Schiff reagent and by its affinity for lectins. Human skin gelatinase shows little or no reactivity toward common protein substrates, such as hemoglobin or casein, and does not cleave helical collagen. Two sites of cleavage in the sequence of gelatin, Gly-Ile and Gly-Leu, have been positively identified using synthetic substrates and tryptic peptides of collagen.
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PMID:Purification and properties of a gelatin-specific neutral protease from human skin. 626 Aug 9

1. Active type collagenase was purified as much as 140-fold from the explant medium of bovine dental sacs and showed a single band on disc gel electrophoresis. Purified collagenase cleaved native collagen at only one locus under physiological conditions, but hydrolyzed neither gelatin nor alpha-casein. The optimal pH was about 7.8. 2. The molecular weight of active type enzyme was 35,000 by gel filtration and 34,000 by gel electrophoresis. The activation of latent type of collagenase resulted in the reduction of molecular weight from 45,000 to 38,000 by gel filtration. 3. A small but detectable amount of collagenase was directly extracted from frozen and thawed bovine dental sacs. In explant media of frozen and thawed tissue and fresh tissue with actinomycin D, some activity was detected for the first 2 days, but essentially no collagenase activity was detected in the explant medium after day 3. 4. The latent type collagenase was activated by trypsin, 4-aminophenylmercuric acetate (4-APMA), thiocyanate and deoxycholate (DOC). DOC showed irreversible dissociation of latent type enzyme in similar fashion to that exerted by 4-APMA. 5. The purified collagenase was inhibited by bovine serum, EDTA, o-phenanthroline, cysteine and dithiothreitol.
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PMID:Purification and characterization of bovine dental sac collagenase. 626 31

Rheumatoid synovial fluid contains an activator of latent collagenase from culture medium of pig synovium. The activator was purified by gel chromatography on Ultrogel AcA 44 and affinity chromatography on soybean trypsin inhibitor coupled to Sepharose 4B. The purified material was homogeneous on SDS-polyacrylamide gel electrophoresis with Mr 88 000. The activator had limited proteolytic activity against azo-casein, but showed amidase activity on Pro-Phe-Arg-NMec, Z-Phe-Arg-NMec, D-Val-Leu-Arg-NPhNO2 and D-Pro-Phe-Arg-NPhNO2, with an optimum at pH 8.0. Activity was completely inhibited by diisopropyl fluorophosphate, soybean trypsin inhibitor, leupeptin and Pro-Phe-Arg-CH2Cl, whereas lima bean trypsin inhibitor, Tos-Lys-CH2Cl, a specific inhibitor of factor XIIa from maize, EDTA and iodoacetate were not inhibitory. These properties of the activator suggested that it might be plasma kallikrein (EC 3.4.21.34), and the possibility was further examined. The activator was treated with [3H]diisopropyl fluorophosphate, and run in SDS-polyacrylamide gel electrophoresis with reduction; a radioautograph of the gel showed a pair of [3H]diisopropyl phosphoryl-labelled bands (Mr 36 000 and 34 000) identical to those obtained with authentic plasma kallikrein. Double immunodiffusion with monospecific antiserum against human plasma kallikrein confirmed the identification. This is the first demonstration of collagenase-activating activity of plasma kallikrein, and raises the possibility that activation of prokallikrein in the inflamed joint space may contribute to the disease process not only by the production of bradykinin, but also by activating latent collagenase.
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PMID:Identification of plasma kallikrein as an activator of latent collagenase in rheumatoid synovial fluid. 627 61

Vibrio vulnificus (lactose-positive vibrio) produced collagenase when grown in 2% synthetic sea salts supplemented with hydrolyzed casein. The addition of collagen or peptone to the medium increased the level of collagenase production. Collagenase activity was inhibited by EDTA but not by fetal calf serum.
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PMID:Collagenolytic activity of Vibrio vulnificus: potential contribution to its invasiveness. 627 15


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