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

1. Homogenates of rat uteri removed 1 and 2 days post partum were centrifuged at 6000 g. Both pellets and supernatants degraded Azocoll, a general proteinase substrate, at pH 7.5. More than 80% of the total activity was in the pellet fraction. 2. Part of the pellet activity was in a latent form. Trypsin and 4-aminophenylmercuric acetate (a thiol-blocking agent) both activated this latent form, indicating that it is an enzyme--inhibitor complex. An endogenous serine proteinase activated part of the latent enzyme during the assay. 3. The enzyme activity was low before parturition and after involution; it was highest during the first 2 days post partum, when the largest losses of uterine wet weight and matrix macromolecules occur. 4. Up to 70% of the enzyme in the pellets was extracted by heating at 60 degrees C for 4 min in 0.1 M-CaCl2/0.05 M-Tris/HCl, pH 7.5. Approx. 30% of the extracted enzyme was still latent. 5. The extracted enzyme was a metalloproteinase, since it was inhibited completely by 1,10-phenanthroline, but not by inhibitors of thiol or serine proteinases. 6. The enzyme was further purified 15--30-fold by gel chromatography and precipitation with (NH4)2SO4. The apparent molecular weight, estimated by gel filtration, was 24000 for the latent form and 12000 for the active form. The pH optimum was 7--7.5. 7. The enzyme also degraded cartilage proteoglycan. This activity was studied by viscometry and the products were analysed by analytical ultracentrifugation. The major product had a mol.wt. of approx. 100000. The sites of cleavage were in the protein core, since no free oligosaccharides were detected. 8. This neutral metalloproteinase is distinct from uterine collagenase and from a uterine metal-dependent endopeptidase that hydrolyses a heptapeptide related to collagen.
 ...
PMID:The extraction of a neutral metalloproteinase from the involuting rat uterus, and its action on cartilage proteoglycan. 701 17

Ultrastructural observations on stigmas in rat ovarian follicles have been performed by scanning electron microscopy. Stigmas were classified into two types under a dissecting microscope; extensively bulging vesicles (bleb-type stigmas) and small, flat avascular areas (flat-type stigmas). The bleb-type stigma had lost its surface epithelial cells extensively, and the flattened and densely arranged fibroblasts without fibrous structures were exposed. These fibroblasts had short, serrate, cytoplasmic projections where multivesicular structure-like granules were seen. By contrast, on the flat-type stigma a small region of exposed stroma was covered with fibrous structures. Removal of the fibrous structures by the HCl-collagenase method revealed that the exposed stroma consisted of stellate fibroblasts surrounding a small opening through which a few granulosa cells were about to discharge. When the cumulus mass was protruding, it was surrounded by the fibrous structures. These findings indicate that both the stellate fibroblast formation and the presence of fibrous structures are needed for the release of ova through the stigma.
 ...
PMID:Scanning electron microscopic studies on stigmas in rat ovaries. 712 59

Aortae from three patients with classic presentation of Marfan syndrome, who died of vascular complications, were subjected to biochemical analyses of the connective tissue; for comparison, aortae from eight age-matched controls, without evidence of connective tissue abnormalities, were examined. Elastin was prepared from the aortae by two techniques. First, the tissues were extracted with 5 M guanidine-HCl, bacterial collagenase digestion and reduction with dithiothreitol (elastin I preparation). Secondly, this material was further purified by extraction with 0.1 M NaOH at 99 degrees C (elastin II preparation). Amino acid analyses of both elastin preparations indicated that the values for desmosine and isodesmosine were reduced in Marfan cases to approximately one-half of the control values. A corresponding increase in lysyl residues was noted in elastin II preparations. Also, the concentration of elastin per milligram dry weight of tissue was reduced in Marfan cases. The hydroxyproline content of elastin was increased in two cases with the Marfan syndrome. Recoveries indicated that the alkali treatment solubilized 46.2% of the elastin I preparations in Marfan aortae compared with 23.7% in controls. In contrast to elastin, the concentration and solubility of collagen were unchanged; the amino acid composition and the genetic types of insoluble collagen isolated by limited pepsin proteolysis were the same in both Marfan and control aortae. The results of our study demonstrate that the cross-linking of aortic elastin is reduced in the three patients with Marfan syndrome. Thus, a defect in elastin could explain the vascular fragility observed clinically in these patients.
 ...
PMID:Marfan syndrome. Demonstration of abnormal elastin in aorta. 717 92

Amyloidosis was induced in mice by the simultaneous injection of sodium caseinate and heat-killed M. butyricum. Amyloid fibrils were isolated by collagenase digestion, 1 M NaCl extraction and repeated washing with 0.15 M NaCl. The amyloid fibril fraction was practically free of contaminations such as collagen, chromatin and membranes as judged by electron microscopic morphometry. The protein AA was purified from the isolated fibrils to an apparent homogeneity as judged by sodium dodecyl sulphate polyacrylamide gel electrophoresis using one step gel filtration from Sephacryl S-200 in the presence of 6 M guanidine-HCl and 50 mM dithiothreitol. The molecular weight of the peptides of the protein AA were 8,500 and 10,000.
 ...
PMID:Purification of amyloid fibrils and protein AA from mouse amyloid deposits induced by caseinate and M. butyricum. 723 20

The effect of surface roughness on osteoblast proliferation, differentiation, and protein synthesis was examined. Human osteoblast-like cells (MG63) were cultured on titanium (Ti) disks that had been prepared by one of five different treatment regimens. All disks were pretreated with hydrofluroic acid-nitric acid and washed (PT). PT disks were also: washed, and then electropolished (EP); fine sandblasted, etched with HCl and H2SO4, and washed (FA); coarse sandblasted, etched with HCl and H2SO4, and washed (CA); or Ti plasma-sprayed (TPS). Standard tissue culture plastic was used as a control. Surface topography and profile were evaluated by brightfield and darkfield microscopy, cold field emission scanning electron microscopy, and laser confocal microscopy, while chemical composition was mapped using energy dispersion X-ray analysis and elemental distribution determined using Auger electron spectroscopy. The effect of surface roughness on the cells was evaluated by measuring cell number, [3H]thymidine incorporation into DNA, alkaline phosphatase specific activity, [3H]uridine incorporation into RNA, [3H]proline incorporation into collagenase digestible protein (CDP) and noncollagenase-digestible protein (NCP), and [35S]sulfate incorporation into proteoglycan. Based on surface analysis, the five different Ti surfaces were ranked in order of smoothest to roughest: EP, PT, FA, CA, and TPS. A TiO2 layer was found on all surfaces that ranged in thickness from 100 A in the smoothest group to 300 A in the roughest. When compared to confluent cultures of cells on plastic, the number of cells was reduced on the TPS surfaces and increased on the EP surfaces, while the number of cells on the other surfaces was equivalent to plastic. [3H]Thymidine incorporation was inversely related to surface roughness. Alkaline phosphatase specific activity in isolated cells was found to decrease with increasing surface roughness, except for those cells cultured on CA. In contrast, enzyme activity in the cell layer was only decreased in cultures grown on FA- and TPS-treated surfaces. A direct correlation between surface roughness and RNA and CDP production was found. Surface roughness had no apparent effect on NCP production. Proteoglycan synthesis by the cells was inhibited on all the surfaces studied, with the largest inhibition observed in the CA and EP groups. These results demonstrate that surface roughness alters osteoblast proliferation, differentiation, and matrix production in vitro. The results also suggest that implant surface roughness may play a role in determining phenotypic expression of cells in vivo.
 ...
PMID:Effect of titanium surface roughness on proliferation, differentiation, and protein synthesis of human osteoblast-like cells (MG63). 754 45

Investigations were carried-out on corneas of rabbit eyes burned with 1N HCl and then treated with low temperature. It was found that cryotherapy has advantageous influence on collagenase activity. In early period after burn cryotherapy could prevent collagenolysis and later inhibited collagenase activity.
 ...
PMID:[Influence of cryotherapy in the inhibition of collagenase activity in experimental corneal burns by hydrochloric acid. Doctoral thesis summary]. 771 56

Experiments were performed to define the best isolation method for isolating Chrysaora fishing tentacle nematocyst organelles in order to minimize non-nematocyst contaminating proteins and proteases and stabilize crude nematocyst venom lethal activity. Techniques employed to disrupt the tentacles included autolysis, homogenization, or digestion using either trypsin or collagenase. Sephacryl-200 gel-filtration chromatography separated two lethal fractions. An immobilized serine protease inhibitor column, m-aminophenyl boronic acid acrylic beads, which reversibly bound one of the two lethal factors, was used in the second and third purification steps. By this means, a 105,000 mol wt. protein was purified, as judged by silver stained SDS-polyacrylamide gels. Lethal activity was inhibited by exposure to the serine protease inhibitor, L-1 chloro 3[4-tosylamido]-7-amino-2-heptanone-HCl, after purification. Although this lethal factor has some characteristics of a serine protease, it is not proteolytically active.
 ...
PMID:Sea nettle (Chrysaora quinquecirrha) lethal factor: purification by recycling on m-aminophenyl boronic acid acrylic beads. 791 86

Quantitative analysis of proteoglycans (PGs) revealed that the content of PG material from cryopreserved aorta, measured as uronate-positive material, was similar to that from fresh tissue (440 +/- 30 versus 430 +/- 7 micrograms/g wet tissue). Gel permeation column chromatography studies suggested that three PG fractions from cryopreserved tissue had molecular weights similar to PG fractions from fresh tissue; K(av) = 0.13, 0.47 (I), 0.20 (II), and 0.43 (III) from cryopreserved tissue and K(av) = 0.13, 0.50 (I), 0.23 (II), and 0.40 (III) from fresh tissue. Sequential extraction of tissue with guanidine-HCl (Gdn-HCl) followed by digestions with collagenase, elastase, and papain also demonstrated that there was no difference between fresh and cryopreserved tissues in the distribution of PGs in the extracts. Transmission electron microscopy analysis revealed less densely packed collagen fibers in cryopreserved tissues compared to fresh tissues. These studies indicate that there is no significant alteration in the content, molecular size, or distribution of PGs in properly cryopreserved tissue.
 ...
PMID:Proteoglycan content in fresh and cryopreserved porcine aortic tissue. 800 93

The cytoarchitecture of the superior cervical ganglion of the common tree shrew was investigated by scanning electron microscopy using the vascular cast technique in conjunction with digestion by collagenase-hyaluronidase/HCl. The main cellular constituents were found to be multipolar neurons that were densely distributed throughout the ganglion. These neurons were covered with a smooth cytoplasmic sheath of satellite cells. After the removal of this sheath by digestion of increased duration, the blebs or knobs on the neuronal surfaces became evident. A meshwork of nerve fibers over the surface of neurons was also observed. Preganglionic sympathetic nerve fibers, giving rise to fine branches that ran toward the postganglionic sympathetic neurons before forming synapses, were demonstrated. Groups of neurons surrounded by capillary loops were also frequently observed.
 ...
PMID:Cytoarchitecture of the common tree shrew (Tupaia glis) superior cervical ganglion. A scanning electron microscope study on vascular cast/enzyme-digested superior cervical ganglia. 811 34

A bone morphogenetic protein purification method for minor quantities of bone material was developed based on collagenase splitting of bone connective tissue. Our aim was to remove and characterize the osteoinductive protein preparation in native form without using strongly dissociative agents. We started from 80 g of HCl-demineralized reindeer bone material which was treated with type I collagen splitting collagenase. The solution was dialyzed against 10 mM glycine-HCl buffer, pH 5.2. The formed precipitate was found to be osteoinductive. After fractionation of the material using HPLC gel filtration it was observed that the high-molecular-weight component of the precipitate was biologically active. Isoelectric focusing revealed that the component consisted of at least eight different protein molecules. Lower-molecular-weight components induced no bone formation. These preliminary findings suggest that in native form at least one part of BMP is in a complex form and other extracellular matrix components bound to the osteoinductive protein complex are significant for BMP action and may act synergistically or as carriers for the BMP.
 ...
PMID:High yield of osteoinductivity can be derived from demineralized bone matrix using collagenase digestion. 815 34


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