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)

A metalloproteinase similar or identical to stromelysin was shown to co-purify with interstitial collagenase from the rat mammary carcinoma cell line, BC1. The mixture of BC1 metalloproteinase and collagenase degraded casein, gelatin, fibronectin, fibrinogen, laminin, proteoglycan and type IV collagen, in addition to types I and II collagen. Using SDS-PAGE and zymography, the Mr of both enzymes was 51.10(3). During storage, the 51.10(3) protein converted to fragments of Mr 34.10(3) and 24.10(3), and isoelectric points of 4.6-5.3 and 5.7-6.0, respectively. The fragments were separated from the intact (Mr 51.10(3) enzymes by DEAE-Sepharose chromatography, but intact metalloproteinase and collagenase activities resisted separation by a range of chromatographic methods. The Mr 34.10(3) fragment retained the proteinolytic activities of the intact enzymes, excepting collagenase cleavage of collagen types I and II. The Mr 24.10(3) fragment had no proteinolytic activity, showed an increase in Mr of 6.10(3) upon reduction, in common with the intact enzymes, and also had similar chromatographic properties to the intact enzymes. The data presented are consistent with a pattern of breakdown which is common to both collagenase and the metalloproteinase, and suggest that both enzymes are comprised of two protein domains.
...
PMID:Identification of a metalloproteinase co-purifying with rat tumour collagenase and the characteristics of fragments of both enzymes. 253 40

Chick-derived native cartilage collagen type X and the pepsin-resistant 45 kDa fragment were susceptible to attack by human synovial collagenase and neutrophil elastase at 25 degrees C and 35 degrees C. Synovial collagenase cleaved type X collagen at two sites which were equally susceptible to the enzyme. In contrast, elastase produced three cleavages, but the sensitive loci showed different susceptibilities as judged by the sequential appearance of specific breakdown products. Both enzymes produced a major, enzyme-resistant fragment of approximately 32 kDa at 35 degrees C, and both of these end-products co-migrated in SDS polyacrylamide gels. Human chondrocyte-derived collagenase also degraded native, 59 kDa collagen type X in a similar manner to that shown by the synovial collagenase. From amino acid sequence data the enzyme cleavages probably occur at three regions of sequence imperfection. The specific cleavages brought about by synovial or chondrocyte collagenase, or neutrophil elastase, may have a functional catabolic role in vivo, and in vitro might provide useful tools with which to further analyse specific properties of the native collagen type X molecule.
...
PMID:Cleavage of collagen type X by human synovial collagenase and neutrophil elastase. 254 40

Human osteoblast cultures (hOB) were examined for the production of interstitial collagenase, tissue inhibitor of metalloproteinases (TIMP), and gelatinolytic enzymes. Cells were isolated by bacterial collagenase digestion of trabecular bone (vertebra, rib, tibia, and femur) from 11 subjects (neonatal to adult). Confluent cultures were exposed to phorbol 12-myristate 13-acetate, PTH, PGE2, epidermal growth factor, 1,25(OH)2 vitamin D3, recombinant human IL-1 beta, and dexamethasone. Collagenase and TIMP were assayed immunologically and also by measurements of functional activity. Collagenase was not secreted in significant quantities by human bone cells under any tested condition. Furthermore, collagenase mRNA could not be detected in hOB. However, hOB spontaneously secreted large amounts of TIMP for at least 72 h in culture. hOB TIMP was found to be identical to human fibroblast TIMP by double immunodiffusion, metabolic labeling and immunoprecipitation, Northern blot analysis, and stoichiometry of collagenase inhibition. SDS-substrate gel electrophoresis of hOB-conditioned media revealed a prominent band of gelatinolytic activity at 68 kD, and specific polyclonal antisera established its identity with the major gelatinolytic protease of human fibroblasts. Abundant secretion of gelatinolytic, but not collagenolytic, enzymes by hOB may indicate that human osteoblasts do not initiate and direct the cleavage of osteoid collagen on the bone surface, but may participate in the preparation of the bone surface for osteoclast attachment by removal of denatured collagen peptides. The constitutive secretion of TIMP may function to regulate metalloproteinase activity.
...
PMID:Human osteoblasts in vitro secrete tissue inhibitor of metalloproteinases and gelatinase but not interstitial collagenase as major cellular products. 254 36

The effect of human TNF on cultured human microvascular endothelial (HME) cells was examined. Incubation with TNF alone transformed the morphology of HME cells from a cobblestone-like appearance into a disordered array of criss-crossed, elongated, spindle-shaped cells. Coadministration of epidermal growth factor (EGF) and TNF caused even more dramatic morphologic changes than TNF alone. Addition of basic fibroblast growth factor or insulin-like growth factor-I showed rather weak effects on cell morphology than EGF. Cell growth of HME cells was stimulated up to two-fold by TNF whereas addition of EGF additively enhanced the growth rate. Treatment of HME cells with 10 ng/ml EGF increased the binding of 125I-TNF, and Scatchard analysis showed increased TNF-R number by EGF treatment. Cellular response to TNF in the absence or presence of EGF was assessed by analyzing SDS-PAGE patterns of secreted proteins from HME cells. TNF enhanced the secretion of a protein of molecular weight 25,000 Da (25 kDa) which was found to be IL-6. In contrast, secretion of a polypeptide of 29 kDa was significantly increased when HME cells were treated with EGF, but not with TNF. Coadministration of TNF and EGF synergistically increased the secretion of the 29-kDa protein. This 29-kDa protein was found to be tissue inhibitor of metalloproteinases when assayed with antitissue inhibitor of metalloproteinases antibody. TNF and EGF also enhanced secretion of collagenase with Mr of approximately 55 kDa. Increased steady state levels of the inhibitor mRNA were observed when HME cells were treated with EGF, and coadministration of TNF further increased the levels. The morphologic transformation of HME cells by TNF and/or EGF is discussed in relation to their expression of the secreted proteins.
...
PMID:Effects of tumor necrosis factor and epidermal growth factor on cell morphology, cell surface receptors, and the production of tissue inhibitor of metalloproteinases and IL-6 in human microvascular endothelial cells. 254 71

The effect of gamma irradiation on the physicochemical properties of injectable human amnion collagen was investigated. Pepsin-extracted human amnion collagen was purified, reconstituted, and irradiated with varying doses of gamma irradiation (0.25 Mrads to 2.5 Mrads). Gamma irradiation had a significant impact on the physical characteristics of the collagen. The neutral solubility of collagen in PBS at 45 degrees C was decreased from 100% for the nonirradiated control sample to 16% for the 2.5 Mrads irradiated sample. SDS polyacrylamide gel electrophoresis also demonstrated the dose-dependent effect of gamma irradiation on collagen cross-links. Electron microscopic observation revealed that even at low irradiation dose (0.25 Mrads), collagen fibril diameter increased. The average diameter was 50 nm for nonirradiated control fibrils, while 4.4 percent of the irradiated collagen fibrils had a diameter greater than 100 nm. Irradiated collagen showed little evidence of damage. Well-preserved cross-striations were found in collagen fibrils at all doses of irradiation. Native amnion collagen irradiated with gamma rays demonstrated a slight increase in resistance to collagenase degradation compared with nonirradiated native collagen samples. Increased resistance to collagenase did not correlate with increasing irradiation dose. After 30 min of incubation at 37 degrees C, both irradiated and nonirradiated collagen was completely digested by collagenase. However, gamma-irradiated collagen did become more sensitive to hydrolysis by trypsin. The higher the irradiation doses used, the greater sensitivity to trypsin was observed. At 0.25 Mrads irradiation only a slight increase was found. No marked differences in amino acid composition were noted among the high dose irradiated, low dose irradiated and control amnion collagen.
...
PMID:The effect of gamma irradiation on injectable human amnion collagen. 255 Apr 67

The role of enzymatic collagen degradation in prostaglandin-induced and physiological cervical ripening was studied in guinea pigs. The cervices were removed from (a) 8 non-pregnant guinea pigs, (b) 8 animals at day 45 of pregnancy, (c) 14 pregnant animals of comparable gestational age which had either an intracervical application of 0.2 ml 5% tylose or 10 micrograms sulprostone gel, and (d) 8 guinea pigs at day 63 to 65 of pregnancy. Collagenase activity was assayed in a highly specific and sensitive system using native collagen type I as substrate. Protease activity was measured by the method of Green and Shaw. Collagen fragments were identified by SDS-polyacrylamide electrophoresis (SDS-PAGE) of acetic-soluble fractions. Collagenase and protease activities were found in all extracts from the different groups. However, there were no differences in enzymatic activities between the non-pregnant, early-pregnant and late-pregnant cervical specimens. Prostaglandin pre-treatment of the cervix led to no significant increase in either collagenase or protease activity as compared to the control groups. The absence of typical collagen degradation products in the SDS-PAGE suggested that no significant collagen breakdown had taken place. In contrast to previously published literature, we conclude that enzymatic collagen degradation is unlikely to be a key factor in prostaglandin-induced and physiological cervical ripening.
...
PMID:Enzymatic collagen degradation in the pregnant guinea pig cervix during physiological maturation of the cervix and after local application of prostaglandins. 255 49

Mast cells have been implicated in the pathogenesis of the matrix degradation observed in the cartilaginous and osseous structures of the rheumatoid joint. We previously reported that human mast cell tryptase, a 134-kD granule-associated neutral protease, is present in rheumatoid synovium and can activate collagenase in crude culture medium in vitro. the present study attempts to depict the precise mechanism of this activation. To express full activation of latent collagenase, matrix metalloproteinase 3 (MMP-3) or stromelysin, can be activated by tryptase in a time and dose-dependent manner. Tryptase was not capable of generating active collagenase in the crude media from cultured rheumatoid synoviocytes depleted of proMMP-3 by immunoadsorption. In addition, the function of the tissue inhibitor of metalloproteinases (TIMP) was not altered by tryptase, and SDS-PAGE analysis revealed no degradation of TIMP by tryptase. The tryptase dependent activation of synoviocyte procollagenase thereby appears to be entirely dependent upon its ability to activate proMMP-3.
...
PMID:Synovial procollagenase activation by human mast cell tryptase dependence upon matrix metalloproteinase 3 activation. 255 80

The secretion of elevated levels of proteinases is considered to be a distinct property of most transformed cells. The cellular and secreted levels of plasminogen activators and collagenases have been examined in the nonmalignant human osteosarcoma (HOS), the malignant Kirsten murine sarcoma virus transformed (KHOS/NP), the temperature sensitive revertant of virus transformed HOS (KHOS-240S) and N-methyl-N'-nitro-N-nitrosoguanidine transformed HOS (MNNG/HOS) clones. Virus and MNNG transformed clones exhibit 100- and 7-fold higher cellular and and 270- and 30-fold higher extracellular plasminogen activator (PA) activity as compared with untransformed HOS controls. The cellular PA activity of the revertant clone is similar to but the secreted level is slightly higher than the HOS controls. SDS-PAGE in the presence of casein and plasminogen is consistent with the major PA species of urinary type (u-PA) and with the absence of PA inhibitor in the parent and revertant clones. The cellular levels of active collagenase are low in all the clones. However, on activation by trypsin, the two active collagenase bands of similar intensity are observed for all the lines in SDS-PAGE in the presence of gelatin. While there appears to be some elevation of secreted collagenase prior to trypsin activation, the activated collagenases appear to have the same size and activity in all of the clones.
...
PMID:Synthesis and secretion of plasminogen activators and collagenases in human cells transformed by Kirsten murine sarcoma virus and N-methyl-N'-nitro-N-nitrosoguanidine. 256 62

Bovine articular and tracheal chondrocytes were cultured at high density in multilayers. Intact or fragmented large aggregating proteoglycans (PG-LA) from cartilage were added to the cultures and the biosynthetic response studied by the incorporation of [3H]-leucine and [35S]-sulfate for proteins and proteoglycans respectively. Incorporated radiolabel and patterns of synthesized macromolecules were compared with control cultures without additives and cultures containing either of the synthetic polymers dextran or dextran sulfate. All proteoglycans and derivatives containing globular protein structures had a stimulatory effect on the biosynthesis of both proteins and proteoglycans as did the highly polyanionic polymer dextran sulfate. Distribution of the radiolabeled material between the cell- and medium pools were however different in the various cultures. A radiolabeled protein, migrating as a triplet band at a position of approximately 140 kDa after reduction, was detected by SDS-PAGE and fluorography. The protein was present in all cell extracts and in the media of cultures stimulated with proteoglycans and proteoglycan fragments, except chondroitin sulfate side chains. The protein was shown to be collagenous in nature by collagenase digestion and identified as procollagen II by immunoprecipitation.
...
PMID:Large cartilage proteoglycan (PG-LA) influences the biosynthesis of macromolecules by isolated chondrocytes. 261 94

A high molecular weight extracellular protein has been purified from cell culture medium of Ewing's sarcoma cell lines, by high performance liquid chromatography and electroelution from SDS-PAGE electrophoresis. This protein has an apparent molecular mass of about 500,000 Da on SDS-PAGE. Immunoprecipitation studies with several extracellular matrix glycoproteins (laminin, fibronectin) specific antisera indicate it is a separate protein. Reduction of disulphide bonds with 2-ME or DTT fails to significantly alter its migration on SDS-PAGE gels, other than a slight apparent increase in molecular mass, indicating an apparent single polypeptide chain structure. The slightly greater mobility observed in unreduced gels suggests one or more regions of intrachain disulfide bonding. It is sensitive to pepsin and trypsin, but resistant to bacterial collagenase indicating that it does not contain collagenous domains. Metabolic labelling with 3H-proline, 3H-leucine, and 35S-methionine indicate that this protein is proline-poor, but leucine, and especially methionine, rich. Sodium 35S-sulfate incorporation is totally negative and treatment with glycosaminoglycan degrading enzymes has no effect on the mobility of the protein on gels, unlike typical proteoglycans. This protein appears by rotary shadowing electron microscopy as a long, thin, filamentous molecule at least 500 nm (0.5 um) in length. The tissue localization and function are unknown at this time, but are under active investigation.
...
PMID:A novel 500,000 Da, linear, single chain extracellular protein synthesized by several childhood tumors. 263 60


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

---