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)

A 140 000 D glycoprotein (140 kD gp), labelled radioactively with surface-specific techniques, remained as the major cell surface glycoprotein in the detergent-resistant cytoskeletal preparations of cultured human fibroblasts. The 140 kD gp was present also in trypsinized cells and was not affected by treatment of the cells either with collagenase, chymotrypsin or thrombin. In density gradient fractionation of whole cells the 140 kD gp was recovered in the plasma membrane fraction together with small amounts of cytoskeletal components. In fractionation of cytoskeletal preparations, on the other hand, the 140 kD gp could not be dissociated from cytoskeletal proteins and together with vimentin it formed the major component of the oligomeric polypeptide complex generated by treating the surface-labelled cytoskeletal preparations with bifunctional cross-linking reagent, dithiobis succinimidyl propionate (DTPS). Moreover, the 140 kD gp seemed to copurify with vimentin upon reconstitution of intermediate filaments from urea-solubilized cytoskeletal preparations. On the other hand, low ionic-induced degradation of vimentin led to a decrease in the amount of the detergent-resistant 140 kD gp on the cell surface. In electron microscopy, a close apposition between bilayer-like plasma membrane remnants of the adherent cytoskeletons and cytoskeletal elements could be seen. The results indicate that the 140 kD gp is a plasma membrane glycoprotein which closely interacts with the detergent-resistant cytoskeleton of cultured human fibroblast. Possible mechanisms of the association are discussed.
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PMID:140 000 Dalton surface glycoprotein. A plasma membrane component of the detergent-resistant cytoskeletal preparations of cultured human fibroblasts. 633 55

The extracellular matrix of cultured chicken embryo fibroblasts undergoes a number of modifications during the early stages of oncogenic transformation. One alteration is increased production of a small protein (Mr approximately 21,000) which is transiently deposited in the matrix by transforming cells infected with LA24, a temperature-sensitive mutant of Rous sarcoma virus (RSV) (Blenis, J., and Hawkes, S.P. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 770-774). This protein is a major component of substratum-associated material (material which remains attached to culture dishes after removal of cells with ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid). Its synthesis is stimulated by transformation of cells with NY68, another ts mutant of RSV, and also by treatment of normal, uninfected cells with the tumor promoter, phorbol myristate acetate. Accessibility of the 21-kDa protein to lactoperoxidase-catalyzed iodination indicates an exposed location within the matrix. The protein binds strongly to the culture dish and/or other matrix components. This interaction can be disrupted by sodium dodecyl sulfate but not by several nonionic detergents, unless beta-mercaptoethanol or KCl (0.5 M) are also present. High concentrations of urea or guanidine hydrochloride also remove the protein from the matrix. The 21-kDa protein is resistant to trypsin, collagenase, and the hydrolytic enzymes associated with cells transformed by the wild-type Prague A RSV but not to Pronase or chymotrypsin. A 21-kDa protein with properties similar to those described above is also detected in the medium and binds to the matrix, suggesting that a potential route of deposition of the 21-kDa protein in the matrix may be via shedding and subsequent interaction with other matrix components.
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PMID:Characterization of a transformation-sensitive protein in the extracellular matrix of chicken embryo fibroblasts. 643 99

A three step extraction procedure was carried out on intact hamster molar tooth germs in vitro labelled with 32PO4 and/or 3H-proline, in order to quantify separately the synthesis of dentine matrix (collagen) and the proline rich enamel matrix proteins. The extraction was based on the high solubility of the proline rich enamel matrix proteins compared with the relatively insoluble dentine matrix collagens. Pretreatment with 10% trichloroacetic acid (step 1) demineralized and removed the non-incorporated amino acids and/or small sized peptides. A consecutive water extraction (step 2) removed a large percentage of the phosphorylated amelogenins as assessed by SDS-urea-polyacrylamide-electrophoresis and amino acid analyses. Collagenase digestibility data showed that only small amounts of collagens were present in this extract. Further extraction with 10% formic acid (step 3) released only small amounts of amelogenins from the explants but also increased contamination with collagens and another predominantly low molecular components. Most of the 3H-activity remaining in the residues was found in the collagenase labile material and was considered to be an appropriate measure for production of dentine collagens. On the other hand, the residues also contained small amounts of 3H-labelled material with the same electrophoretic mobility as amelogenins but had much more 32P-activity than the amelogenins derived from the water and formic acid extracts. It is suggested that this material in the residues probably contains the crystal bound enamel matrix proteins.
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PMID:Biosynthesis of tooth germ proteins in vitro: a fast quantitative extraction of amelogenins from intact hamster molar tooth germs. 659 32

The globular domain of collagen IV was solubilized by collagenase digestion from a mouse tumor, human placenta and bovine aorta and was purified by chromatographic methods. The materials show a unique, mainly non-collagenous amino acid composition and contain small amounts of glucosamine and galactosamine. The globular structures with Mr = 170 000 appear as a hexameric assembly originating from two collagen IV molecules. Subunits of this assembly are two different dimers Da and Db (Mr about 56 000) and monomers (Mr = 28 000). Their N-terminal amino acid sequences start with short triple-helical sequences, which overlap with the C-terminal triple helix of the alpha 1(IV) and alpha 2(IV) chain, demonstrating that the globule originates from the C terminus of collagen IV. Dimers arise from monomers by disulfide cross-linking (form Db) and/or formation of non-reducible cross-links (form Da). Reduction under non-denaturing conditions causes partial dissociation of the globule and of collagen IV dimers, indicating that reducible cross-links are formed between monomers of two different collagen IV molecules. Dissociation of the hexamer into the subunits can be achieved with 8 M urea, sodium dodecyl sulfate or in the pH range 2.5-4. The latter indicates that carboxyl groups are essential for association. Mixtures of the subunits (monomers and dimers) or purified dimers reassemble in neutral buffer into hexamers as shown by ultracentrifugation and electron microscopy. Reconstituted hexamers, however, dissociate in a much broader pH range than the native globules. Circular dichroic spectra indicate that the structure is more completely refolded from acid-treated than from urea-treated material. These data suggest that globules originating from monomers (as existing in single collagen IV molecules) are stabilized by the adjacent triple helix. Covalent cross-link formation stabilizes the globular structure and allows reconstitution in stoichiometric proportions.
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PMID:Subunit structure and assembly of the globular domain of basement-membrane collagen type IV. 669 21

A rabbit antiserum raised against ACI rat liver biomatrix was used to identify components common to biomatrix and plasma membranes of adult hepatocytes. Biomatrix was isolated from intact rat livers by reverse perfusion via the inferior vena cava with sodium deoxycholate, nucleases and lipid extracting solvents. Immunoprecipitation analysis of detergent extracts of hepatocytes surface-labeled with 125I indicated that antibodies, purified from anti- biomatrix antiserum by adsorption and desorption from intact hepatocytes, showed reactivity with a single MW 105 kD component, designated Hep 105. Indirect immunofluorescence analysis showed that Hep 105 was expressed in some regions of the perisinusoidal space and in all three domains of the hepatocyte plasma membrane and was present on some but not all of the fibrous elements in frozen sections of biomatrix . The presence of Hep 105 on biomatrix was confirmed by immunoprecipitation analysis which showed that Hep 105 was present in components solubilized from biomatrix by sequential treatment with 0.5 M acetic acid, 0.05% collagenase and 4 M urea. Further characterization using immunoprecipitation analysis in combination with immobilized lectins and two-dimensional polyacrylamide gel electrophoresis (PAGE) indicated that Hep 105 was a non-collagen glycoprotein which showed charge heterogeneity and existed on the cell surface as a disulfide-linked heterodimer of apparent MW 125 kD. Two hybridomas, constructed by fusing P3 X 63Ag8 myeloma cells with spleen cells from mice immunized with intact hepatocytes, were shown by immunodepletion and two-dimensional gel electrophoretic analysis to be secreting monoclonal antibodies (Mab) against Hep 105. Examination of frozen sections of rat liver stained by indirect immunofluorescence showed that reactivity of both Mabs was concentrated in the bile canalicular domain of the hepatocyte plasma membrane, suggesting that the reactive epitopes were not accessible in the sinusoidal and intercellular membrane domains. Taken together, these results suggest that Hep 105 may play a role in the interactions between hepatocytes and extracellular matrix.
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PMID:Cell surface expression by adult rat hepatocytes of a non-collagen glycoprotein present in rat liver biomatrix. 672 95

After extraction of proteoglycans and soluble matrix proteins from canine puppy rib cartilage, with 4 M guanidine . HCl, the insoluble collagen-containing residue has been shown to contain two collagenase-resistant structural glycoproteins, A and G. The characteristic subunits of these insoluble structural glycoproteins have been identified by solubilizing them with 50 mM dithiothreitol (DTT) in 1% sodium dodecyl sulfate (SDS) and 8 M urea and comparing the SDS disc gel patterns with those of more readily soluble matrix proteins. Three subunit bands which did not occur in gels from the soluble matrix proteins were found in the solubilized material from both the collagen-containing residue and the structural glycoproteins. One band, of about 87 000 daltons, was found equally in both A and G glycoproteins. The other two bands formed a closely spaced doublet, of about 30 000 and 27 500 daltons, of which the lower band is present in higher concentration in G. Although none of these SDS gel bands corresponds with the 220 000 dalton band produced by pure fibronectin under the same conditions, a fraction of the solubilized glycoprotein material resembles fibronectin in showing an affinity for collagen, fibrinogen, heparin, and an antibody to plasma fibronectin. Crude fibronectin from human plasma contains minor components (including one of about 87 000 daltons) which show partial identify in immunodiffusion reactions with components of the solubilized cartilage structural glycoproteins. Solubilized A gave a stronger reaction with anti-plasma fibronectin than did G and the soluble matrix proteins have no reaction. It is possible either that the intercellular structural glycoproteins are formed by selection of some of the partial cleavage products of fibronectin which occur in connective tissues as well as in plasma, or that cleavage products of tissue structural glycoproteins occur in plasma which cross-react with anti-plasma fibronectin.
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PMID:Comparison of cartilage structural glycoproteins with matrix proteins and fibronectin. 678 99

Rabbit aortic intima-media fragments were incubated with [14C]mannose and [3H]fucose for 6 h to detect glycoproteins synthesized in situ. The radioactively labelled and the non-labelled samples were extracted with 0.2 mM-CaCl2/0.5 mM-dithiothreitol/0.5 mM-ATP and chloroform/methanol/water (4:4:1, by vol.). The delipidated residue was extracted with 5 M-guanidinium chloride/0.05 M-dithiothreitol/0.1 M-Tris/0.4% Na2EDTA, pH 7.5, before (extract 1) and after hydrolysis with collagenase (extract 2). The proteins in extracts 1 and 2 were S-carboxamidomethylated and separated by molecular-sieve chromatography, polyacrylamide-gel electrophoresis and isoelectric focusing in sucrose gradients in urea. The apparent molecular weights of glycoproteins were 36 000 (glycoprotein I) from extract 1, 50 000 (glycoprotein II) and 130 000 (glycoprotein III) from extract 2. The molecular weights of the non-labelled and radioactively labelled glycoproteins were identical. Glycoproteins I, II and III contain large amounts of polar amino acids and methionine. They contain neither hydroxyproline nor 3-methylhistidine. A hydroxyproline-containing component of 160 000-apparent-mol.wt. relatively rich in polar amino acids and labelled with incorporated sugars was isolated from extract 1. The incorporation in vitro of radioactive sugars into glycoproteins I, II, III and collagenous glycoproteins indicates that they are synthesized in the surviving aorta by the smooth-muscle cells.
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PMID:Structural glycoproteins from rabbit aortic media. 687 Aug 24

Isolated hepatocytes obtained by collagenase perfusion of adult rat livers were seeded on collagen gels and kept in a chemically defined culture medium (except for the first 6 h of culture where 10% fetal calf serum was added). Cells adopted an epitheloid shape within 4 h and arranged themselves in a trabeculae-like pattern during the first 20 h of culture. In the electron microscope numerous tight junctions and bile capillaries were observed at sites of cell-to-cell contact. From metabolite analyses in the culture medium the following conclusions can be drawn: The cells continued to synthesize urea and ketone bodies for 5 days of culture. The cytosolic and mitochondrial redox states of the nicotinamide adenine nucleotide systsm were as in the liver in vivo and the oxygen supply of hepatocytes was sufficient under the culture conditions. Maximal velocities of ketogenesis from octanoate and of urea formation from ornithine plus ammonium chloride were stable during a 120 h culture period and compared well with rates found in the isolated perfused rat liver.
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PMID:Morphology and metabolism of adult rat hepatocytes in primary culture. 699 70

After extraction of bovine corneal stroma with 1M CaC1(2) and subsequent digestion of the insoluble residue with purified bacterial collagenase, two crude structural glycoprotein (SPG) fractions were obtained; one which precipitated upon dialysis against water of the collagenase solubilized material and the other which was extracted by 8M urea from the collagenase insoluble material. Amino acid analyses of these crude SGP fractions indicated that they were primarily non-collagenous but that very small amounts of collagen-derived amino acids were present. Upon gel filtration of these SGP fractions on Sepharose 4B-CL, void volume fractions were isolated from each of the crude fractions which were enriched, relative to the original crude fractions, in the collagen-derived amino acids. Carbohydrate analysis indicated that the void volume fractions had the properties of glycoproteins rather than proteoglycans. Upon disulfide reduction and SDS-PAGE, each of these fractions was resolved into five major protein bands with molecular weights of 155,000, 137,000, 117,000 82,000 and 34,000. Only the three largest bands contained the collagen derived amino acids. These data are consistent with the presence within bovine corneal stroma of a large structural glycoprotein complex comprised of at least five protein components associated through disulfide bonds. Collagen apparently is associated with three of these, either through covalent crosslinkage, or as part of the primary structure.
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PMID:Isolation and preliminary characterization of a structural glycoprotein complex from bovine corneal stroma. 718 8

Somatic extracts of Nippostrongylus brasiliensis contain protease inhibitor(s) capable of inhibiting the activity of trypsin and chymotrypsin A and B. This inhibitor was partially purified by affinity chromatography. Its molecular weight is in the range of 9500-10 000. The inhibition of both trypsin and chymotrypsin depends on the same or closely adjacent active sites of the inhibitor molecule. The inhibitor is unaffected by heating, pH changes or urea, but is sensitive to 2-mercaptoethanol The formation of the enzyme-inhibitor complex is time-dependent. The complex does not dissociate with KC1. The inhibitor has no effect on the activity of elastase, subtilisin, pepsin, rennin, papain and collagenase.
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PMID:A protease inhibitor of Nippostrongylus brasiliensis. 725 48


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