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 sandwich enzyme immunoassay for collagenase inhibitor was set up with a pair of monoclonal antibodies prepared against bovine dental pulp collagenase inhibitor. Two different combinations of the antibodies were found to be applicable to the immunoassay; one was for the determination of both bovine and human collagenase inhibitors, but the other was only for the bovine one. Minimum sensitivity was 1 pg/tube (3 pg/ml) for bovine collagenase inhibitor and 1.5 pg/tube (5 pg/ml) for human inhibitor. The levels of collagenase inhibitor in several normal human body fluids were measured by the immunoassay, and the values obtained were as follows: serum, 183 +/- 30 ng/ml (mean +/- SD); platelet-poor plasma, 68 +/- 13; cerebrospinal fluid, 60 +/- 13; amniotic fluid, 1,780 +/- 969; tear fluid, 418 +/- 145; mixed saliva, 316 +/- 166; urine, 7 +/- 8.
...
PMID:A sandwich enzyme immunoassay for collagenase inhibitor using monoclonal antibodies. 254 Apr 3

The neutral zinc metalloendopeptidase (NEP, EC 3.4.24.11) is an integral membrane protein found in brain tissue, polymorphonuclear leukocytes, and many epithelia. We show here that NEP is expressed on rabbit synovial fibroblasts and on simian virus 40 (SV40) DNA- and H-ras-transformed rabbit mammary epithelial cells. Treatment of these cells with 12-O-tetradecanoylphorbol-13-acetate (TPA) for 24 h decreased expression of NEP mRNA transcripts and decreased the biosynthetically labeled immunoprecipitable NEP antigen. In contrast to its effects on NEP, TPA treatment induced expression of the secreted metalloproteinase collagenase and the tissue inhibitor of metalloproteinases. TPA induced stromelysin, another secreted metalloproteinase, only in the fibroblasts. These data provide evidence that the expression of the membrane-bound NEP is regulated in several cell types.
...
PMID:Phorbol diesters regulate expression of the membrane neutral metalloendopeptidase (EC 3.4.24.11) in rabbit synovial fibroblasts and mammary epithelial cells. 254 98

The soft tissues adjacent to osseointegrated dental implants (OII) were investigated using clinical, biochemical and microbiological methods. Tooth and implant crevices were compared in 15 partially edentulous patients, examining 28 peri-implant and 19 periodontal sites, and in 6 edentulous patients, examining 13 implant sites. Sites were classified by standard periodontal indices; the crevicular fluid flow determined; crevicular fluid was collected for collagenase assays; and the subgingival bacterial flora was examined and cultured. Differences in clinical parameters were noted in that implants had significantly less keratinized gingiva and deeper probing depths. Crevicular fluid was present in the OII sulcus but the crevicular fluid flow did not differ from that observed from tooth sites either in the partially edentulous or edentulous patients. Tissue collagenase activity and collagenase inhibitor were detected in the implant crevicular fluid and, as in periodontal sites, a strong inverse relationship was found between the levels of active collagenase and collagenase inhibitor. Microbiology included darkfield microscopy, anaerobic culturing for total colony forming unit counts and identification of black pigmented Bacteroides (BPB). Few differences were observed between implants and teeth in partially edentulous patients, indicating that crevices around teeth may act as reservoirs of bacteria which can colonize implant sites. A higher percentage of BPBs and wet spreaders (Capnocytophaga) was noted at partially edentulous implant sites when compared with edentulous implant sites, perhaps reflecting the lower numbers of periodontal pathogens present in edentulous mouths. Overall, the characteristics of implant sulci appear to be similar to periodontal sulci with respect to crevicular fluid flow and microflora.
...
PMID:Microbiota and crevicular fluid collagenase activity in the osseointegrated dental implant sulcus: a comparison of sites in edentulous and partially edentulous patients. 254 14

A new, high molecular weight (66,000 daltons) inhibitor of collagenase (LCI) has been isolated and partially characterized. It accounted for 20% of the collagenase-inhibitory activity in the supernatants of rabbit chondrocytes cultured in 10% acid-treated fetal bovine serum (ATFBS). LCI was stable to 60 degrees C and sensitive to reduction and alkylation. Unlike a low molecular weight collagenase inhibitor, similar to Tissue Inhibitor of Metallo-Proteinases (TIMP), it did not bind to concanavalin A-Sepharose 4B.
...
PMID:A high molecular weight collagenase inhibitor made by rabbit chondrocytes in cell culture. 254 42

A potent collagenase inhibitor was purified from cells of calf aorta medial tissue maintained in culture. This molecule was characterized and identified as TIMP (Tissue Inhibitor of Metalloproteinases). Formation of a TIMP--collagenase complex was demonstrated chromatographically using pure TIMP and pure pig synovial cell collagenase. The N-terminal aminoacid sequence of TIMP was determined and, using appropriate oligonucleotide probes the human genes was cloned from a human cDNA bank. This gene was expressed in E. coli, and fully active TIMP was obtained after a denaturation renaturation process. The interest of TIMP as a model for the design of novel collagenase inhibitors is discussed.
...
PMID:[Natural inhibitor of metalloproteinases: structural and functional study]. 254 70

Tumor cell motility and the passage of tumor cells through various tissue matrices, including basement membrane, are important components of the metastatic process. Proteolytic enzymes, including a type IV collagen-specific collagenase, have been demonstrated to play a significant role in extracellular matrix and basement membrane degradation. In addition, exogenous collagenase has been shown to enhance the motility of some tumor cells independent of its effect on collagen-containing material. Previous studies have also indicated that collagen fragments are chemotactic for many tumor cells. We therefore studied the effect of type I and type IV collagen-specific collagenases, other enzymes involved in collagenase activation and connective tissue degradation, and subsequent collagen degradation products on the directed migration of tumor cells. We report that type I and type IV collagen-specific mammalian collagenases were potent chemoattractants as were native type I and type IV collagens and collagen fragments. Collagenase inhibitor SC44483 inhibited the type IV collagenase-stimulated migration. Collagenase pretreatment of the tumor cells potentiated the migratory response of the tumor cells to collagen and collagen fragments. The plasminogen activator, urokinase, as well as plasminogen itself also enhanced the directed migration of tumor cells in concentrations that suggest involvement of the appropriate cell surface receptor. The chemotactic response of tumor cells to the proteases studied extends the prior report of a role for collagenases and other matrix-active enzymes in tumor cell behavior in addition to matrix degradation.
...
PMID:Directed migration of murine and human tumor cells to collagenases and other proteases. 254 19

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

Pump-1 cDNA has recently been isolated by screening a human tumor cDNA library with a transin (rat stromelysin) probe under low-stringency hybridization conditions. The cDNA codes for a potential protein with significant sequence similarity to the metalloproteinases collagenase and stromelysin, but which lacks the hemopexin-like domain characteristic of these enzymes. Expression of pump-1 cDNA in cos cells using an expression vector leads to secretion of a protein of Mr 28,000 with latent, organomercurial-activatable proteinase activity. Cos cells transfected with a partial pump-1 cDNA in the vector pPROTA secrete a fusion protein between the IgG-binding domains of staphylococcal protein A and pump-1. The fusion protein binds to IgG-Sepharose, and the bound fusion protein undergoes apparent autocleavage in the presence of 4-aminophenylmercuric acetate with elution of active pump-1 species of Mr 21,000 and 19,000. Active pump-1 degrades casein, gelatins of types I, III, IV, and V, and fibronectin and can activate collagenase. Active pump-1 is inhibited by EDTA, 1,10-phenanthroline, and the tissue inhibitor of metalloproteinases. These results show that, despite the absence of a hemopexin-like domain, pump-1 is a latent secreted metalloproteinase. Postpartum rat uteri contain elevated levels of rat pump-1 mRNA. On the basis of this observation, its size, and its substrate specificity, we suggest that pump-1 might correspond to a previously described uterine metalloproteinase, matrix metalloproteinase 7.
...
PMID:Pump-1 cDNA codes for a protein with characteristics similar to those of classical collagenase family members. 255 50

The validity of the enzymatic assay of procollagenase within crude biological media containing also the collagenase inhibitor TIMP (tissue inhibitor of metalloproteinases) as well as other (pro)metalloproteinases and sometimes, metalloproteinase-TIMP complexes, has been reevaluated. To be enzymatically assayed, procollagenase has to be activated. The standard activation procedures by either trypsin or 4-aminophenylmercuric acetate (APMA) both allow an optimal recovery of collagenase from procollagenase when the media do not contain free TIMP. However, they do not destroy TIMP nor do they reactivate the collagenase present in enzyme-inhibitor complexes. Therefore, the collagenase formed by the activation of procollagenase in the presence of free TIMP is immediately inactivated by binding to the inhibitor. As a result, both the bound collagenase and TIMP can no longer be assayed by enzymatic methods. An optimal recovery of collagenase can, however, be obtained if free TIMP is neutralized by the binding of other tissue metalloproteinases (such as those present in culture media of rabbit bone marrow-derived macrophages) prior to the activation and assay of procollagenase. Similarly, it is possible to recover under an active free form a large part of the TIMP present in collagenase- (or other metalloproteinase-)TIMP complexes by heating the complexes at acid pH under conditions which inactivate the collagenase.
...
PMID:The enzymatic evaluation of procollagenase and collagenase inhibitors in crude biological media. 255 82


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

---