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)

Synthesis of collagen types I, II, III, and IV in cells from the embryonic chick cornea was studied using specific antibodies and immunofluorescence. Synthesis of radioactively labeled collagen types I and III was followed by fluorographic detection of cyanogen bromide peptides on polyacrylamide slab gels and by carboxymethylcellulose chromatography followed by disc gel electrophoresis. Type III collagen had been detected previously by indirect immunofluorescence in the corneal epithelial cells at Hamburger-Hamilton stages 20--30 but not in the stroma at any age. Intact corneas from embryos older than stage 30 contain and synthesize type I collagen but no detectable type III collagen. However, whole stromata subjected to collagenase treatment and scraping (to remove epithelium and endothelium) and stromal fibroblasts from such corneas inoculated in vitro begin synthesis of type III collagen within a few hours while continuing to synthesize type I collagen. As demonstrated by double-antibody staining, most corneal fibroblasts contain collagen types I and III simultaneously. Collagen type III was identified biochemically in cell layers and media after chromatography on carboxymethylcellulose be detection of disulfide-linked alpha l (III)3 by SDS gel electrophoresis. The conditions under which the corneal fibroblasts gain the ability to synthesize type III collagen are the same as those under which they lose the ability to synthesize the specific proteoglycan of the cornea: the presence of corneal-type keratan sulfate.
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PMID:Synthesis of type III collagen by fibroblasts from the embryonic chick cornea. 624 16

Plasminogen is present in the cornea andcan be activated to plasmin by plasminogen activator. Plasmin is able, in turn, to activate latent collagenase. This system could initiate and perpetuate the collagen degradation of corneal ulceration. This report details evidence for such a system in the cornea. Plasmin has been found to activate latent collagenase from organ cultures of ulcerating rabbit corneas and from fibroblast cultures derived from such corneas. As in the case of activation by trypsin, activation by plasmin results in the conversion of the 40,000 MW latent form to an active species of 23,000 MW. Explants of normal or alkali-burned, ulcerating corneas demonstrated plasminogen-dependent lysis of fibrin clots; frozen sections of such corneas demonstrated that lysis begins in the superficial stroma near the periphery of the cornea. Multiply freeze-thawed ulcerating corneas, but not normal corneas, showed initial lysis, not peripherally but at the ulcer region containing polymorphonuclear leukocytes. The fact that the peripheral lytic pattern existed in corneas that were obtained from eyes prefrozen in liquid nitrogen before excision of the corneas would suggest that plasminogen activator is normally contained in cells in vivo and is not made only in response to tissue injury. There was no correlation between the location of blood vessels or the presence of the corneal endothelium and the plasminogen-dependent lysis. Plasminogen activator from the ulcerating cornea and from fibroblasts was characterized by sodium dodecyl sulfate--gel electrophoresis of its cleavage products of plasminogen. The activator cleaves plasminogen into heavy- and light-chain fragments similar to those produced from plasminogen by urokinase. Plasminogen activator activity was quantitated by a new assay that restricts diffusion of the enzyme to one dimension into a narrow bore tube. The addition of plasminogen daily to cultures of ulcerating corneas resulted in earlier rises of plasminogen activator, collagenase, and collagen degradation fragments in the culture media. Although total plasminogen activator levels were not increased by the addition of plasminogen to culture, levels of both collagenase and solubilized collagen were approximately doubled. It is concluded that the plasminogen activator--plasmin system might play an important role in the destruction of stromal matrix in corneal ulceration.
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PMID:Evidence for a role of the plasminogen activator--plasmin system in corneal ulceration. 625 12

Media conditioned by epithelial cells from the adult rabbit cornea were capable of both stimulating and inhibiting production of latent collagenase by stromal cells from the same source. Cytochalasin B was required in this in vitro system for both secretion of stimulators by epithelial cells and production of collagenase by stromal cells in response. Optimal production of collagenase by stromal cells in response. Optimal production of stimulators occurred in low-density epithelial cell cultures. Chromatographed conditioned medium from such cultures contained three stimulator fractions with apparent molecular weights of 19,000, 54,000, and greater than or equal to 90,000. High-density epithelial cell cultures secreted inhibitors of stromal cell collagenase production with apparent molecular weights of 7000 and 19,000. Cytochalasin B was not required for production of inhibitors. Inhibitory conditioned medium blocked the effect of the 19,000-dalton and 54,000-dalton stimulator on stromal cells. The data suggest that epithelial cells, in ways depending on their density, may modulate collagen degradation in the integument.
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PMID:Regulation of stromal cell collagenase production in adult rabbit cornea: in vitro stimulation and inhibition by epithelial cell products. 625 75

Medroxyprogesterone, dexamethasone, or cortisone, locally applied in sustained release polymer to rabbit V2 carcinoma implanted in the rabbit cornea, blocked neovascularization and three-dimensional growth of the tumor. These hormones similarly prevented the vascular proliferative response to implants in the rabbit cornea of mouse B-16 melanoma and also the response to implants of polymer containing tumor extract with angiogenesis activity. The inhibitory responses were accompanied by considerable reduction in collagenolytic activity released into culture medium by explants of the two tumors and of the corneal region containing angiogenic hepatoma extract. Morphologic studies revealed extensive three-dimensional disruption of the compact laminated collagenous structure of the cornea by untreated V2 carcinoma. In the presence of hormone the tumor grew slowly as a noninvasive two-dimensional plaque limited to the narrow region of the insertion pocket in the cornea, with no obvious disturbance of structure elsewhere. Cortisone was much les effective than medroxyprogesterone or dexamethasone. Testosterone and estradiol had no effect on the three measured properties. The data suggest that local hormonal interference with neovascularization, collagenase production, and tumor growth can prevent neoplastic invasion and destruction of a dense collagenous connective tissue.
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PMID:Inhibition of tumor growth, vascularization, and collagenolysis in the rabbit cornea by medroxyprogesterone. 626 56

In previous studies we found that a mild thermal burn of the vitamin A-deficient rat cornea caused collagenase release into the medium of corneas placed in culture media 72 hr after applying the burn. Collagenase was released only on day 1 of culture and was not released from identically burned corneas of control rats. We now demonstrate that in deficient corneas, this collagenase release on day 1 of culture increased gradually with increasing time between burn and sacrifice, reaching a maximum at 16 hr after burning a remaining high up to 72 hr. In control rats day-1 collagenase release also increased to a maximum at 16 hr after the burn but then declined to almost zero at 72 hr. Trypsin treatment of day-1 media from both control and deficient corneas, taken at 72 hr after the burn, showed an almost complete absence of latent (inhibited) collagenase. Histologic observations revealed a close correlation between the presence of infiltrating polymorphonuclear neutrophils (PMNs) and the ulcerative lesions seen in burned, deficient corneas. When PMN infiltration was blocked by application of a tissue adhesive, no ulceration occurred and collagenase activity in the day-1 media dropped to almost zero. If burned and unburned areas of deficient corneas were separated and cultured separately, the burned area (containing most of the PMNs) was found to have 10 times the collagenase activity of the unburned area. In the controls, PMNs and some collagenase activity was detectable only 16 hr after the burn. We concluded that in the burned, vitamin A-deficient cornea there is increased attraction of PMNs to the lesion, resulting in collagenase release by these and possibly other cells, and ultimately resulting in ulceration.
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PMID:Studies on the source and release of collagenase in thermally burned corneas of vitamin A-deficient and control rats. 627 20

Analysis of cell populations in the cornea may be performed rapidly and accurately employing the technique of enzymatic disaggregation. To illustrate this method normal rat corneas and corneas infected 24 and 48 hours previously with Staphylococcus aureus were disaggregated in a solution containing pancreatin and collagenase. The cells released were counted and identified morphologically. These results were compared to cell counts made from histologic sections. Over 95% of the corneal cells were viable after the disaggregation and leukocytes obtained from the infected corneas retained their phagocytic capacity. This approach allows sensitive analysis of cell populations in a wide range of corneal conditions, including infection and allograft rejection.
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PMID:Enzymatic disaggregation of the infected rat cornea. 629 40

We have examined the ability of primary adult rabbit skin cells to regulate collagenase production in vitro. Dermal cells constitutively produce collagenase in culture, and enzyme production by these cells can be influenced by epithelial cells. Co-culture with skin epidermal cells resulted in more enzyme production by dermal cells, whereas co-culture with corneal epithelial cells yielded less enzyme activity. Connective tissue cells from a different source, cornea, also produced collagenase when co-cultured with skin epidermal cells, although the stromal cells alone made no enzyme. The drug cytochalasin B had very little influence on collagenase production by dermal cells, either alone or in co-culture with epidermal cells, but did significantly potentiate enzyme production by corneal stromal cells responding to epidermal effector molecules. Epidermal-cell-conditioned medium from both fetal and adult rabbit skin was a potent source of stimulators (apparent mol wt 20,500 and 55,000) of connective-tissue-cell collagenase production. Stimulator production by epidermal cultures was cell density dependent. Optimal production of stimulators occurred in adult cultures containing 10(6) epidermal cells/ml of medium, and in fetal cultures containing 10(5) cells/ml. Inhibitors of connective tissue cell enzyme production were not detected in conditioned medium from either adult or fetal epidermal cells.
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PMID:Regulation of connective tissue collagenase production: stimulators from adult and fetal epidermal cells. 632 86

Polymorphonuclear leukocytes (PMN) are important in corneal disease because of their role as effector cells in inflammation and ulceration. The favorable effect of citrate on corneal ulceration appears to result from inhibition of the PMN. Citrate does not enter the cells but chelates Ca2+ in the extracellular fluid and may promote a loss of some intracellular Ca2+. Isocitrate is the only tricarboxylic acid cycle intermediate that inhibits PMN, also by Ca2+ chelation. When isobutylcyanoacrylate is polymerized, a substance, probably formaldehyde, inhibitory to PMN, continuously leeches from the plastic. Although acetylcysteine has been reported to inhibit collagenase in vitro it has a direct effect of enhancing the respiratory burst and possibly degranulation of PMN stimulated by opsonized zymosan. Dexamethasone had no effect on PMN stimulation while prednisolone was partially inhibitory at high concentrations. Indomethacin exerts an inhibitory effect on all parameters of PMN stimulation. These studies clarify the site and mechanism of citrate action as well as show the importance of knowing the effect of drugs on the PMN.
Cornea
PMID:The effect of citrate and other compounds on PMN incubated in vitro: further studies on the site and mechanism of action of citrate. 657 89

The structural alterations elicited in the rabbit corneal stroma by experimental Serratia marcescens keratitis and by a highly purified serratia protease preparation were compared by gross observation, biochemical analyses, and electron microscopic examination of the affected tissue. Acute inflammation, liquefactive necrosis of the cornea, and descemetocele formation occurred during the development of the infection and after the intracorneal injection of submicrogram amounts of the protease. In vitro incubation of insoluble corneal stromal tissue with the bacterium or with the protease resulted in solubilization of the stromal proteoglycan ground substance; however, specific collagenase activity was not detected. Electron microscopic examination of corneas damaged by the bacterial infection and by the protease revealed loss of ruthenium red staining of the proteoglycan ground substance and dispersal of ultrastructurally normal collagen fibrils. Thus, our findings indicate that the major corneal damage which occurs during serratia keratitis and after the injection of the serratia protease is caused by solubilization and loss of the ground substance of the tissue. In addition, the observation that the major structural alterations observed during serratia keratitis can be reproduced by the bacterial protease supports the idea that the enzyme is involved, at least in part, with the production of severe corneal damage by the bacterium.
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PMID:Characterization of rabbit corneal damage produced by Serratia keratitis and by a serratia protease. 702 49

We measured the relative solubility of collagen in acetic acid after pepsin digestion and tentatively identified the types of collagen present in corneas of rabbits of various ages and in corneal scar tissue, using hydroxyproline assays and polyacrylamide gel electrophoretic analyses. More than 80% of the collagen in normal developing rabbit cornea was soluble after pepsin treatment; no more than 45% of that in two-week-old corneal scars was soluble. The predominant collagens in normal cornea and healing tissue were types I and AB. Type AB increased from 6% of the total collagen in fetal cornea to 11% in cornea from young adults. Collagen from two-week-old corneal wounds contained 16% type AB. Corneal type AB collagen was less soluble and more resistant to degradation by mammalian collagenase than was type I collagen. Unlike the normal cornea, in healing tissue the relative rate of synthesis of type I to type AB collagens did not correspond to their deposition. These results suggest a basic alteration in the molecular structure of the corneal scar, which may be instrumental in preventing the healing tissue from producing a normal, functioning organ.
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PMID:Quantitative analysis of collagen from normal developing corneas and corneal scars. 729 90


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