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

---

Query: EC:3.4.24.3 (collagenase)
18,340 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Single smooth muscle cells were isolated from circular muscle of the canine gastric corpus by collagenase incubation. Cytoplasmic pH (pHi) of these cells was measured fluorometrically using the trapped dye 2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein. Cells were examined for their Na+/H+ exchange activity after intracellular acidification. Cells acid-loaded by propionate exposure, the NH4+ prepulse technique or suspension in a Na+-depleted medium regained almost normal pHi upon exposure to a Na+ medium. The Na+-dependent alkalinization was amiloride sensitive. As well, addition of amiloride to cells suspended in a Na+ medium caused a concurrent decrease in pHi. The study indicates that a Na+/H+ antiport is present in these smooth muscle cells.
...
PMID:Evidence for a Na+/H+ antiport in stomach smooth muscle cells. 283 89

On treatment with collagenase, brain microvessels, together with several protein components, lose some enzymatic activities such as alkaline phosphatase and gamma-glutamyltranspeptidase, whereas no change occurs in the activities of 5'-nucleotidase and glutamine synthetase. The energy-requiring "A-system" of polar neutral amino acid transport is also severely inactivated, whereas the L-system for the facilitated exchange of branched chain and aromatic amino acids is preserved. In the collagenase-digested microvessels, this leads to loss of the transtimulation effect of glutamine on the transport of large neutral amino acids, because such transtimulation is due to a cooperation between the A- and L-systems. By contrast, NH4+ maintains (and even enhances) its ability to stimulate the L-system of amino acid transport, presumably through glutamine synthesis within the endothelial cells.
...
PMID:Isolated brain microvessels as in vitro equivalents of the blood-brain barrier: selective removal by collagenase of the A-system of neutral amino acid transport. 289 Jul 11

Human skin, maintained in serum-free organ culture, secretes a neutral metalloendopeptidase which is remarkably specific for gelatin. Because the product peptides from the action of collagenase on collagen become denatured into random coil polypeptides of 25000 and 75000 daltons at physiological temperature, it is thought that this "gelatinase" is the second, and possibly the only other enzyme in the pathway of extracellular collagen degradation. New types of high-performance liquid chromatography (HPLC) columns have enabled us to improve the yields of active gelatinase from skin culture medium. Raw medium, which has been dialyzed and lyophilized, is fractionated with ammonium sulfate, and applied to Pharmacia Blue Sepharose in a batch step. The 0.4 M sodium chloride eluate is then subjected to gel filtration on Sephacryl S-200, followed by gradient elution from Amicon Green Sepharose. The fractions with gelatinolytic activity are applied to a Bio-Rad TSK-Phenyl-5PW HPLC column for mild hydrophobic chromatography with a gradient of decreasing ammonium sulfate concentration. In the final step, the enzyme is applied to a Pharmacia Mono-Q FPLC column and eluted with a gradient of sodium chloride. At this point, the enzyme appears as two bands, corresponding to enzymatic activity zymograms on sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
...
PMID:Purification of gelatin-specific neutral protease from human skin by conventional and high-performance liquid chromatography. 299 26

Collagen types synthesized by murine bone marrow cells were studied and the effect of lithium chloride on collagen biosynthesis in vitro was investigated. In the liquid culture system used, an adherent, mixed cell population supports hemopoiesis. Radioactive labeling of cell cultures and subsequent fractionation with ammonium sulfate, enzyme digestion, immune precipitation, and gel electrophoresis indicated that the bone marrow cells synthesized precursors to collagen types I, III, and IV, and fibronectin. A previously undescribed molecule or fragment with an apparent molecular weight of 17,000 daltons that was susceptible to bacterial collagenase and containing no interchain disulfide bonds was also identified in the culture media of both control and lithium-treated cells. Lithium treatment did not affect the types of collagen synthesized, although the relative proportions of collagen types may differ from controls. However, lithium does have an effect on the appearance of some, as yet unidentified, non-collagenous components in the cell culture media.
...
PMID:Collagen synthesis by murine bone marrow cell culture. 301 14

Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis of pepsin-solubilized collagens from post-burn granulation tissues revealed that type V collagen consisted of 3 alpha chains: alpha 1(V), alpha 2(V), and alpha 3(V). The mean value (0.12 +/- 0.01 SD) of the type V/type I ratio in the granulation tissues was significantly higher (p less than 0.001) than that (0.03 +/- 0.01 SD) of the ratio in normal skin. The average ratio of alpha 1(V):alpha 2(V):alpha 3(V) of type V collagen purified from the granulation tissues was determined to be about 5:3:1. SDS-polyacrylamide gel electrophoresis patterns of 3 alpha chains were not affected in the presence or absence of 2-mercaptoethanol. Purified type V collagen was degraded by bacterial collagenase, but remained intact after tadpole collagenase digestion, in contrast to type I and type III collagens. Amino acid analyses of each alpha chain separated on SDS-gel electrophoresis of type V collagen revealed that all 3 alpha chains of type V collagen were poor in alanine, rich in hydroxylysine, and had high ratios of hydroxylysine/lysine, which are typical features of type V collagen. The purified type V collagen was further fractionated by ammonium sulfate into 2 molecular species, [alpha 1(V)]2 alpha 2(V) and alpha 1(V)alpha 2(V)alpha 3(V). Our data demonstrate that type V collagen in preparations from human post-burn granulation tissues consists of 3 alpha chains and can be resolved into 2 distinct heterotrimers.
...
PMID:Isolation and characterization of type V collagen from human post-burn granulation tissues. 302 Jan 32

The purpose of this study has been to compare collagen-gelatin degrading enzymes isolated from cancer cell organelles and cytosol to the metalloproteinases released by cancer cells. To this end, metastatic mouse melanoma cell organelles were isolated by sucrose density gradient centrifugation and metalloproteinases were assayed using native and denatured [methyl-3H]collagen substrates. Solubilized proteinases were purified by ammonium sulfate precipitation, anion exchange, concanavalin A affinity and gel-filtration column chromatographic procedures and characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The conclusions were as follows: malignant melanoma cells have a metalloproteinase (Mr = 59,000) which is shed from cells into conditioned medium as a component of intact membrane vesicles rather than as a soluble enzyme; storage of tumor-conditioned medium leads to the generation of autoactivated soluble metalloproteinases of lower molecular weight; purification of these metalloproteinase species yielded variant collagenases that have considerable gelatinolytic activity and a cleavage preference site for the Gly-Ile bond in a collagen-like synthetic octapeptide substrate which is typical for collagenase-type metalloproteinases. It is proposed that localization of potent proteinases to the surface of cancer cells facilitates the local breakdown of connective tissues during the invasive process.
...
PMID:Metastatic mouse melanoma cells release collagen-gelatin degrading metalloproteinases as components of shed membrane vesicles. 303 Apr 44

The major collagenolytic proteinase present in the culture filtrate of Bacillus cereus (strain Soc 67, isolated from the human oral cavity) has been purified to homogeneity by a procedure that comprised concentration of ultrafiltered growth medium on a Millipore PTTK00005 membrane, precipitation with ammonium sulfate, gel permeation chromatography, chromatofocusing, fast protein liquid chromatography on an anion-exchange column, and finally fast protein liquid chromatography on a gel column. The enzyme hydrolyzed, with decreasing rates, phenylazobenzyloxy-carbonyl-L-Pro-L-Leu Gly-L-Pro-D-Arg (PZ-PLGPA), furylacrylolyl-L-Leu-Gly-L-Pro-L-Ala, and furylacryloyl-L-Phe-Gly-Gly, while furylacryloyl-Gly-L-Leu-NH2 was not hydrolyzed. The enzyme degraded soluble and insoluble collagens, Azocoll and gelatin. Bradykinin was hydrolyzed at a high rate at the Phe-Ser bond. The enzyme was sensitive to pyrophosphate, L-cysteine, and L-histidine and could be totally inactivated in the presence of metal chelators. The enzyme contains 1 mol of Zn/mol and the hydrolysis of PZ-PLGPA is slightly increased by Ca2+. The enzyme is readily inhibited by heavy metal cations, but Cu2+ and Ni2+ affected the catalysis in opposite ways: increasing levels of Cu2+ decreased the affinity of the enzyme for PZ-PLGPA, whereas Ni2+ had no effect. The effect of Cu2+ also depended on the pH and type of buffer used. Detailed chemical modification experiments suggested that the active site of the enzyme contains at least 1 tyrosyl and 1 lysyl residue, and 1 carboxyl group. The enzyme was not sensitive to sulfhydryl reagents and thiols did not activate the enzyme. The modification studies were unable to reveal active histidyl residues. The ability of the enzyme to hydrolyze PZ-PLGPA, furylacryloyl-L-Leu-Gly-L-Pro-L-Ala, furylacryloyl-L-Phe-Gly-Gly, and various collagenous materials, its inactivity toward furylacryloyl-Gly-L-Leu-NH2, and the results from the chemical modification studies suggest that the B. cereus (Soc 67) collagenolytic enzyme can be regarded as a true collagenase which resembles the Clostridium histolyticum collagenase(s).
...
PMID:Purification and properties of an extracellular collagenolytic protease produced by the human oral bacterium Bacillus cereus (strain Soc 67). 304 Jul 51

Analysis of collagenolytic activity in gingival crevicular fluid (GCF) has revealed the presence of an enzyme capable of fragmenting native 3/4- and 1/4-collagen cleavage products generated by collagenase. An enzyme with similar activity was also identified in media conditioned by fibroblasts from rat periodontal ligament and gingiva, and by rat osteoblastic cells (ROS 17/2.8, 17/2A, 17/2B). In culture, the enzyme was secreted in a latent form that could be activated by organomercurials. For further characterization of this novel enzyme (MMP-V), the osteoblast proteinase was partially purified. ROS 17/2.8 conditioned medium was harvested daily and the 25%-60% sat. ammonium sulfate fraction chromatographed on an AcA 54 gel filtration column. Latent forms of MMP-V (apparent Mr approximately 54 k) and collagenase (Mr approximately 54 k) were resolved from gelatinase (Mr approximately 76 k) and two collagenase inhibitors (Mr approximately 62 k, approximately 36 k). Activated MMP-V degraded native 3/4-collagen fragments from collagen types I and II in a step-wise manner and was active on denatured collagen. MMP-V showed a divalent cation requirement, was active at neutral pH, and was inhibited by collagenase inhibitor and fetal bovine serum, but not by serine, thiol, or carboxyl proteinase inhibitors. These properties indicate that MMP-V is a member of the matrix-degrading, neutral-metalloproteinase family of enzymes which include collagenase, gelatinase, stromelysin, and telopeptidase. The enzyme may function in the degradation of collagen fibrils by cleaving proteinase-resistant 3/4-collagen fragments that are stabilized by association with neighboring collagen molecules.
...
PMID:Initial characterization of a neutral metalloproteinase, active on native 3/4-collagen fragments, synthesized by ROS 17/2.8 osteoblastic cells, periodontal fibroblasts, and identified in gingival crevicular fluid. 304 Aug 31

Urine from monocytic leukemia and other febrile patients contains an inhibitor of interleukin 1 (IL-1), as measured by prostaglandin E2 and collagenase production by human fibroblasts and synovial cells. With the use of recombinant IL-1, the IL-1 inhibitor was partially purified by using ammonium sulfate precipitation, anion-exchange, and gel filtration chromatographies. IL-1 inhibitory activity elutes with an 18,000 to 25,000 apparent molecular size. The same fractions also inhibit IL-1 assayed by the proliferation of murine thymocytes and human fibroblasts. Both forms of human recombinant IL-1, IL-1 alpha and IL-1 beta, which show only 26% homology, but nevertheless bind to the same receptor, are affected by this natural inhibitor to the same extent. In contrast, human recombinant tumor necrosis factor, which shares some of the biologic activities of IL-1, is not inhibited by the urinary IL-1 inhibitor. This study shows that the various biologic activities of both forms of human recombinant IL-1 are inhibited by a partially purified natural urine-derived factor.
...
PMID:A urine inhibitor of interleukin 1 activity affects both interleukin 1 alpha and 1 beta but not tumor necrosis factor alpha. 304 Aug 55

Glycoproteins were extracted from human atherosclerotic lesions by a phosphate buffered heparin solution and by 0.15 M NaCl, followed by sequential digestion of the tissue by collagenase and elastase. Proteins obtained from the extracts by fractional precipitation by (NH4)2SO4 at 40%, 60% and 100% saturation of the salt at pH 4.0 were analyzed for total protein and for fibronectin and laminin by immunodiffusion. Greater amounts of proteins were extracted from atherosclerotic lesions than from uninvolved tissue. The buffered heparin solution extracted several-fold more protein than 0.15 M NaCl. Fibronectin was found in most protein fractions from every extract, but the presence of laminin was noted only in the protein fractions precipitated at 40% saturation of (NH4)2SO4 in the extracts of the tissue by heparin.
...
PMID:Identification of fibronectin and laminin in glycoprotein extracts of human aorta. 393 60


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>

---