Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: 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)

Matrix vesicles are present in the calcifying front and in the site of callus formation of fracture heeling. In calcifying process, matrix vesicles have important roles. The metalloprotease was isolated from matrix vesicles and subsequently characterized. Matrix vesicles obtained from chicken epiphysial cartilage by collagenase digestion and differential centrifugation were further purified by Sepharose CL2B gel filtration. The protease was solubilized from the vesicles and isolated by Sephadex G-150 gel filtration. Disc electrophoresis of the enzyme gave a single protein band. The matrix vesicle protease had a MW of 33,000 daltons, an optimal pH of 7.2, and was inhibited 100% by 0.1 mM EDTA and 0.2 mM o-Phenanthroline. alpha 2-Macroglobulin, ovalbumin, cysteine, penicillamine, ethane-1-hydroxy-1, 1-diphosphonate (EHDP) and pyrophosphate at higher concentrations were also inhibitory. The inhibition by o-phenanthroline was reversed by Co2+, Zn2+, Fe2+ and Cu2+. The protease released from the matrix vesicle at the calcifying front could degrade non-collagenous protein moieties which inhibit precipitation of minerals in the extra-vesicular matrix and thus facilitate mineralization.
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PMID:[Isolation and characterization of a metalloprotease associated with chicken epiphyseal cartilage matrix vesicles]. 309 Jan 76

A small metalloproteinase that digests Azocoll was found in the uterus of the rat. Its activity increased to high levels during the postpartum period in parallel with the breakdown of the extracellular matrix exclusive of collagen (Sellers, A., and Woessner, J.F., Jr. (1980) Biochem. J. 189, 521-531). This enzyme has now been purified almost 7,000-fold to homogeneity from 12 g of tissue using molecular sieve chromatography, blue sepharose chromatography, and zinc-chelate chromatography. Gel electrophoresis with sodium dodecyl sulfate and dithiothreitol gives Mr = 28,000 for the latent form of the enzyme and Mr = 19,000 for the active form that arises spontaneously or by treatment with aminophenylmercuric acetate. The enzyme digests components of the extracellular matrix including gelatins of types I, III, IV, and V, fibronectin, and proteoglycan. It digests the alpha 2(I) chain of gelatin in preference to the alpha 1(I) chain and cleaves dinitrophenyl-Pro-Leu-Gly-Ile-Ala-Gly-Pro-D-Arg. It cleaves the B chain of insulin at two points: Ala14-Leu15 and Tyr16-Leu17. It has no action on collagens of types I, III, IV, or V at 26 degrees C and no action on elastin or phenylazo-Pro-Leu-Gly-Pro-D-Arg. The pH optimum is at pH 7 and the pI at 5.9. The enzyme requires zinc and calcium ions for activity; cobalt and strontium can partially replace these metal ions. The enzyme is not inhibited by low levels of phosphoramidon or Zincov. Its properties clearly distinguish it from collagenase, gelatinase (matrix metalloproteinase 2), and stromelysin (matrix metalloproteinase 3); it therefore constitutes a further member of the family of extracellular matrix metalloendopeptidases. The name matrix metalloproteinase 7 is proposed.
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PMID:Purification and properties of a small latent matrix metalloproteinase of the rat uterus. 318 22

A neutral metalloproteinase has been isolated and purified from adherent rheumatoid synovial cells in culture. This protease, named matrix metalloproteinase 3, (MMP-3) degrades gelatin, proteoglycan, fibronectin, type IV collagen, laminin, and the N propeptide of type I procollagen. It can be separated from MMP-2 (a potent gelatinase), and MMP-1, an interstitial collagenase. MMP-3 is released from cells as a proenzyme of 55 Kda. Activation by trypsin or organic mercurials produces 2 active species of 45 Kda and 28 Kda. The enzyme contains zinc as an intrinsic component and requires calcium for conformational stability. In concert, active MMP-1, -2, and -3 can destroy all significant structural proteins of joint structures.
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PMID:Matrix metalloproteinases 1, 2, and 3 from rheumatoid synovial cells are sufficient to destroy joints. 330 38

Cadmium (Cd) induces testicular tumors of interstitial cell (IC) origin in rats which can be prevented by zinc (Zn). Zn-induced synthesis of metallothionein (MT), a metal-binding protein with a high affinity for Cd, is thought to account for tolerance to Cd in most tissues by sequestration of Cd. However, the mechanism of Zn inhibition of Cd-induced carcinogenesis in the testes is unknown. Our studies with ICs obtained by collagenase dispersion of rat testes, indicate the levels of the Cd-binding protein in ICs are unaltered by Zn. This testicular protein also was found to differ from MT in amino acid content and to have a lower affinity for Cd. Thus, MT does not seem to be involved in protection of ICs against Cd carcinogenesis. Altered Cd toxicokinetics as a possible explanation for Zn-induced tolerance was therefore explored. Cd uptake into isolated ICs had passive diffusion and nonpassive (carrier mediated or active transport or both) components. The nonpassive component of Cd accumulation was markedly reduced by addition of Zn in vitro, indicative of competition for uptake at the cellular level. These results indicate that toxicokinetic alterations leading to reduced Cd accumulation may play an important role in Zn induction of tolerance to Cd carcinogenesis in the testes.
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PMID:In vitro assessment of target cell specificity in cadmium carcinogenesis: interactions of cadmium and zinc with isolated interstitial cells of the rat testes. 339 32

A metalloprotease has been isolated from matrix vesicles of chicken epiphyseal cartilage and subsequently characterized. Matrix vesicles obtained by collagenase digestion and differential centrifugation were further purified by Sepharose CL2B gel filtration. The protease was solubilized from the vesicles by treatment with deoxycholate and freeze-thawing, and then isolated by Sephadex G150 gel filtration. Disc electrophoresis of the enzyme, which displayed protease activity toward azocasein substrate, gave a single protein band. Based on molecular weight (MW) determination, lack of immunocross reactivity, and differences in electrophoretic migration, there is little possibility of any contamination with external protease from the commercial collagenase used for vesicle preparation. The matrix vesicle protease had a MW of 33,000 and a pH optimum of 7.2 and was completely inhibited by 0.1 mM EDTA and 0.2 mM o-phenanthroline. alpha 2-Macroglobulin, ovalbumin, cysteine, penicillamine, ethane-1-hydroxy-1, 1-diphosphonate (EHDP) and pyrophosphate at higher concentrations were also inhibitory. The inhibition by omicron-phenanthroline was reversed by Co2+, Zn2+, Fe2+, and Cu2+. Protease activity was most abundant in the heavy fraction of matrix vesicles fractionated by discontinuous sucrose density gradient centrifugation. Release of this protease at the calcifying front could degrade noncollagenous protein moieties that inhibit precipitation of minerals in the extravesicular matrix and thus facilitate mineralization.
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PMID:Isolation and characterization of a metalloprotease associated with chicken epiphyseal cartilage matrix vesicles. 352 86

Previous studies have shown that the administration of cadmium causes extensive necrosis of the testes and, eventually, a high incidence of interstitial cell tumors. However, the interactions of cadmium with interstitial cells of the testes have not been well defined. Therefore, this study was designed to assess the uptake of cadmium into this potential target cell of cadmium carcinogenesis. Interstitial cells were prepared by collagenase dispersion of decapsulated Wistar rat testes and separated from seminiferous tubules by unit gravity sedimentation. Such preparations showed a high exclusion rate of trypan blue. The interstitial cell preparations were incubated at 33 degrees with various concentrations of cadmium (1.0 to 100 microM) for periods ranging from 0.5 to 60 min. At the end of the incubation, cellular cadmium was separated from cadmium in the media by centrifugation through an oil layer. Initial experiments showed three distinct phases of cadmium influx into interstitial cells, a primary rapid velocity phase (V0; 0 to 1.5 min), a second intermediate velocity phase (V1; 3 to 12 min), and a third low velocity phase (V2; 15 to 60 min). V2 appeared to have both influx and efflux components, as efflux experiments indicated an approximate 20% loss of cadmium from 15 to 60 min. The initial phase was found to be nonsaturable and was not decreased by inclusion of potassium cyanide (1.0 mM), N-ethylmaleimide (1.0 mM), or zinc (100 microM) in the incubation mixture. However, V1 was found to be saturable between 50 and 100 microM cadmium and was substantially decreased by the inclusion of potassium cyanide, N-ethylmaleimide or zinc during incubation. These data suggest that cadmium is taken up into interstitial cells by a transport system that may normally function in zinc uptake and may possibly constitute carrier mediated or active transport.
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PMID:Interactions of cadmium with interstitial tissue of the rat testes. Uptake of cadmium by isolated interstitial cells. 401 91

The six collagenases (alpha, beta, gamma, delta, epsilon, and zeta) from Clostridium histolyticum isolated in the preceding paper [Bond, M. D., & Van Wart, H. E. (1984) Biochemistry (first paper of three in this issue)] have been characterized in detail. The molecular weights determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis range from 68 000 to 125 000. Isoelectric focusing experiments demonstrate that the isoelectric points of the collagenases are in the 5.35-6.20 range. These experiments also reveal that the subspecies of alpha- and gamma-collagenases (alpha1 vs. alpha 2 and gamma 1 vs. gamma 2) have different isoelectric points but the same molecular weights. Microheterogeneity is also observed for the beta- and epsilon-collagenases. The amino acid compositions of all six collagenases have been determined, and analysis for neutral sugars and hexosamines shows that none of the enzymes have a significant carbohydrate content. Zinc and calcium are the only metals that copurify with the collagenases. The purified enzymes contain approximately 1 mol of zinc/mol of protein and a calcium content that varies from about 2 mol/mol for alpha-collagenase to about 7 mol/mol for beta-collagenase. All of the collagenases are 5-10 times more active against gelatin than collagen. The alpha-, beta-, and gamma-collagenases are significantly less active toward the synthetic peptide substrates examined than the delta-, epsilon, and zeta-collagenases. This property, taken together with data on the stabilities and amino acid compositions of these enzymes, strongly supports their assignment to two distinct classes. This establishes clearly that C. histolyticum does, indeed, produce more than one different type of collagenase.
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PMID:Characterization of the individual collagenases from Clostridium histolyticum. 608 88

Two siblings with junctional epidermolysis bullosa are described: both survived beyond parturition. They were treated with the usual therapeutic doses of phenytoin, dapsone, prednisolone, and zinc supplements without effect. Investigation of the skin of one of the patients showed that his fibroblasts, collagen synthesis and collagenase levels were normal. In view of the normality of the collagenase levels, it is probably not surprising that phenytoin was ineffective. Electron microscopy demonstrated junctional cleavage without pathology in the dermis itself: abnormal hemidesmosomes were seen as described previously, though it is suggested that this is not the primary abnormality which results in the disease process.
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PMID:Junctional epidermolysis bullosa in two siblings: clinical observations, collagen studies and electron microscopy. 609 44

Three collagenases from Clostridium histolyticum, designated C1, C2, and C3, with apparent molecular weights of 96 000, 92 000, and 76 000 were purified. Peptide maps of the enzymes prepared by digestion with Staphylococcus aureus V-8 protease were found to be similar. Cleavage of native C1 with alpha-chymotrypsin or V-8 protease yielded C2 and C3. This suggested that proteolysis of the Mr 96 000 collagenase may have occurred in vivo, producing the other two lower molecular weight enzymes. Previously prepared antiserum directed against a form of the bacterial enzyme similar by molecular weight and charge to collagenase C3 and Fab' fragments generated from this antiserum inhibited the collagenolytic activity. C1, C2, and C3 were immunologically identical by Ouchterlony double diffusion, and C3 was able to compete with C1 for the antiserum binding site. The ability of each enzyme to bind to antiserum raised against the bacterial collagenase supported the hypothesis that these three proteins were closely related. Zinc analyses of C1 and C3 resulted in a value of 1.14 mol of zinc/mol of C1 and 0.82 mol of zinc/mol of C3. C1 did not contain carbohydrate as measured by gas-liquid chromatography or periodic acid-Schiff staining.
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PMID:Purification and characterization of three forms of collagenase from Clostridium histolyticum. 609 92

ATP pyrophosphohydrolase was partially purified from fetal bovine epiphyseal cartilage. The purification was about 10- and 100-fold over the enzyme activities of matrix vesicle fraction and cell homogenate, respectively. The pyrophosphohydrolase and alkaline phosphatase were separated by a sequential application of Sepharose CL-6B and DEAE-cellulose column chromatographies. The purified enzyme migrated as a single band corresponding to the molecular weight of 230,000 in sodium dodecyl sulfate-polyacrylamide disc gel by electrophoresis. The enzyme absolutely required Zn2+ for its activity and appeared to bind Zn2+ strongly with an apparent affinity of p[Zn2+]0.5 = 13.4. The apparent Km for ATP was 0.18 mM. The enzyme was also reactive toward various nucleoside triphosphates including GTP, CTP, and UTP. In contrast, various phosphodiesters including RNA, UDP-glucose, NAD, and bis-p-nitrophenylphosphate were 5% or less as reactive as the nucleoside triphosphates. The pyrophosphohydrolase was inactive toward adenosine 3':5'-monophosphate or various phosphonates. UDP-glucose (1 mM), NAD (1 mM), or RNA (1 mg/ml) failed to inhibit the ATP pyrophosphohydrolase activity. These observations suggest that the ATP pyrophosphohydrolase of the cartilage is probably not a phosphodiesterase I. The matrix vesicle fraction, which probably also included some plasma membrane vesiculated during collagenase digestion, contained the highest specific activity of the enzyme as compared to other subcellular fractions of either epiphyseal or articular cartilage.
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PMID:Purification and partial characterization of ATP pyrophosphohydrolase from fetal bovine epiphyseal cartilage. 621 90


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