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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)
1. A method is described for the isolation of rat parotid acinar cells by controlled digestion of the gland with
trypsin
followed by
collagenase
. As judged by Trypan Blue exclusion, electron microscopy, water, electrolyte and ATP concentrations and release of amylase and lactate dehydrogenase, the cells are morphologically and functionally intact. 2. A method was developed for perifusion of acinar cells by embedding them in Sephadex G-10. Release of amylase was stimulated by adrenaline (0.1-10muM), isoproternol (1 or 10 MUM), phenylephrine (1 muM), carbamoylcholine (0.1 or 1 muM), dibutyryl cycle AMP (2 MM), 3-isobutyl-1-methylxanthine (1mM) and ionophore A23187. The effects of phenylephrine, carbamoylcholine and ionophore A23187 required extracellular Ca2+, whereas the effects of adrenaline and isoproterenol did not. 3. The incorporation of 45Ca into parotid cells showed a rapidly equilibrating pool (1-2 min) corresponding to 15% of total Ca2+ and a slowly equilibrating pool (greater than 3h) of probably a similar dimension. Cholinergic and alpha-adrenergic effectors and ionophore A23187 and 2,4-dinitrophenol increased the rate of incorporation of 45Ca into a slowly equilibrating pool, whereas beta-adrenergic effectors and dibutyryl cyclic AMP were inactive. 4. The efflux of 45Ca from cells into Ca2+-free medium was inhibited by phenylephrine and carbamoylcholine and accelerated by isoproterenol, adrenaline (beta-adrenergic effect), dibutyryl cyclic AMP and ionophore A23187. 5. A method was developed for the measurement of exchangeable 45Ca in mitochondria in parotid pieces. Incorporation of 45Ca into mitochondria was decreased by isoproterenol, dibutyryl cyclic AMP or 2,4-dinitrophenol, increased by adrenaline, and not changed significantly by phenylephrine or carbamoylcholine. Release of 45Ca from mitochondria in parotid pieced incubated in a Ca2+-free medium was increased by isoproterenol, adrenaline, dibutyryl cyclic AMP or 2,4-dinitrophenol and unaffected by phenylephrine or carbamoylcholine. 6. These findings are compatible with a role for Ca2+ as a mediator of amylase-secretory responses in rat parotid acinar cells, but no definite conclusions about its role can be drawn in the absence of knowledge of the molecular mechanisms involved, their location, and free Ca2+ concentration in appropriate cell compartment(s).
...
PMID:Calcium metabolism and amylase release in rat parotid acinar cells. 18 53
A new immunochemical procedure was introduced to estimate the nephritogenic activity of
collagenase
and pronase digests of various rat organs. A glycoprotein isolated from
collagenase
digests of various rat organs showed the nephritogenic activity as well as the antigenic activity that induces nephrotoxic antibody, which were nearly identical to those of a glycoprotein isolated from
trypsin
digests of the rat organ concerned. A glycoprotein isolated from pronase digests of various rat organs was proved to have no antigenic activity that induces nephrotoxic antibody, but the existence of nephritogenic activity was proved in this glycoprotein. These results were supported firmly by the assay experiments for nephritogenicity. Ouchterlony gel diffusion test and the assay experiments demonstrated the existence of a common nephritogenic substance among the glycoproteins isolated from
trypsin
,
collagenase
and pronase digests of rat organs.
...
PMID:Nephritogenic glycoprotein. V. Immunochemical studies on nephritogenic activity of collagenase and pronase-digests prepared from various rat organs. 19 62
1. The involuting rat uterus displays an extremely rapid breakdown of collagen. Collagenase activity can be assayed directly in the insoluble 6000g pellet of uterine homogenates. At 1 day post partum, about 85% of this
collagenase
activity is in a latent form. 2. This latent form can be activated by
trypsin
or by a serine proteinase present in the uterine pellets. 3. The activating enzyme of the tissue is inhibited by a wide spectrum of
trypsin
inhibitors, including Trasylol, soya-bean and lima-bean
trypsin
inhibitors, snail inhibitor and di-isopropyl phosphoro-fluoridate. Partial inhibition is produced by benzamidine, phenylmethanesulphonyl fluoride, epsilon-aminohexanoate, leupeptin, antipain and alpha1-antitrypsin. Ovomucoid, 7-amino-1-chloro-3-tosylamido-1-heptan-2-one and 1-chloro-4-phenyl-3-(N-benzyloxy-carbonyl)amino-L-butan-2-one are not inhibitory. 4. Extraction of uterine pellets with 0.1 M-CaCl2 at 60 degrees C releases both latent and active
collagenase
. Exclusion chromatography on Sephadex G-100 gives an apparent molecular weight of approx. 77000 for the latent form and 66000 for the active form. The latent form is suggested to be a zymogen of
collagenase
.
...
PMID:A latent form of collagenase in the involuting rat uterus and its activation by a serine proteinase. 19 99
Human skin procollagenase has been isolated, in pure form, from the medium of fibroblasts cultured in the presence or absence of added serum. Purification was achieved using a combination of cation-exchange (phosphocellulose or carboxymethylcellulose) and gel-filtration chromatography. Two forms (60 000 and 55 000 daltons) of the procollagenase were detected by electrophoresis in sodium dodecyl sulfatepolyacrylamide gels and could be separated by chromatography on Ultrogel AcA-44. Each form was converted to active enzyme by
trypsin
, producing species of 50 000 and 45 000 daltons, respectively. An autoactivation process also occurred, which yielded active enzyme without a detectable change in molecular weight. Procollagenase also was found in organ cultures of human skin but only when serum was added to the medium. This suggests that a serum-inhibitable proteolytic system is present in these cultures which, like
trypsin
, converts procollagenase to the active enzyme forms that can be isolated from serum-free organ culture medium. The
collagenase
species obtained from either fibroblast or organ culture medium were chromatographically and electrophoretically identical.
...
PMID:Human skin collagenase: isolation of precursor and active forms from both fibroblast and organ cultures. 19 68
The authors present morphological characteristics of primary monolayer cultures prepared from the pancreas of bovine fetuses. Combined treatment with
trypsin
and collalytine solutions (a preparation with
collagenase
activity) was used for dispersion of the tissue of the pancreas. Numerous epithelial cells corresponding by morphofunctional characteristics to beta-cells of the islets of Langerhans were contained in be cultures obtained; an aldehyde-fuchsin-positive granularity was revealed in the cytoplasm of these cells. Degranulation of these cells occurred under the effect of an increased glucose concentration in the nutrient medium.
...
PMID:[Cell cultures obtained using collalytin from the pancreases of cattle fetuses]. 19 89
The composition of extracellular proteinases from Actinomyces fradiae 0072 and their effect on proteins and synthetic substrates were studied. The enzyme preparation was found to have keratinolytic, caseinolytic, collagenolytic,
collagenase
,
trypsin
-like, carboxy- and aminopeptidase activities. Five low molecular weight proteinases capable to hydrolyse keratin, casein, azocollagen were obtained via fractionation of the enzyme preparation on DEAE-cellulose and Sephadex columns. Proteinases from Act. fradiae and aminopeptidase with a molecular weight of 31,000 were shown to be different enzymes. In hydrolysis of L-leucyl-2-naphthylamide the Michaelis constant and Vmax of the enzyme were found to be 3.02 X 10(-3) M and 0.35 X 10(2) micronM/ml-min, respectively.
...
PMID:[Characteristics of extracellular proteolytic enzymes of Actinomyces fradiae 0072]. 19 35
In a qualitative and quantitative study of enzymic dispersion of cells from the mucosal layer stripped from canine urinary bladder,
trypsin
was found to be equal or superior to the other enzymes tested for dispersal of urothelial cells specifically. Collagenase or
collagenase
plus
trypsin
served to disperse the whole tissue. A procedure for recovering the urothelial cells as a single-cell suspension and establishing them in culture is presented. The morphology, culture behaviour, and chromosome complement of these cells is described.
...
PMID:Canine urinary bladder epithelial cells: preparation for cell culture by enzyme dispersion. 19 76
Although it has been shown that keratome-sliced skin contains active adenylate cyclase systems which respond to various hormones and drugs, unequivocal proof that the epidermis contains these hormone-responsive systems is still lacking. We demonstrate in this study that "pure" epidermis obtained after either
collagenase
or
trypsin
treatment does contain the hormone-sensitive adenylate cyclase systems.
...
PMID:Epidermal adenylate cyclase systems: the retention of hormone responsiveness after enzymatic separation of pure epidermis. 19 27
Serum-free media of minced tissue cultures of VX-2 rabbit carcinoma contained a specific collagenolytic activity capable of releasing soluble radioactive peptides from [14C]-labeled collagen fibrils. It was also capable of reducing the viscosity of acid-soluble collagen solutions by cleaving the tropocollagen (TC) molecules primarily at one site to TCA (75%) and TCB (25%) fragments. Three chromatographic fractions were separated by gel filtration: F1, (MW 85-110,000) present in larger amounts in early cultures of younger tumor tissue; F2, (MW-35-40,000) the major component with maximum production in the day 3 media of younger and advanced tumor tissues; F3, (MW 18-22,000) the minor component. Early cultures of younger tumor tissue contained a latent
collagenase
and were subject to
trypsin
activation suggesting the presence of inactive enzyme precursors or an enzyme-inhibitor complex.
...
PMID:Changes in the collagenolytic activity released by primary VX-2 carcinoma cultures as a function of tumor growth. 19 82
Samples of 35 tumors from the head and neck region (25 squamous cell, 2 basal cell, 5 parotid, 3 melanoma, and 1 lymphosarcoma) were cultured after dispersement with either
trypsin
or
collagenase
treatment. Growth was established in 14 (40%). Cultured tumor cells were then used as target cells in in vitro assays of patients' cellular and humoral immunity to their own or similar tumors. Preliminary data suggest this may be a reliable method of monitoring responses in patients receiving immunotherapy for head and neck malignancies.
...
PMID:Tissue-cultured head and neck tumors: their use in in vitro assays of immune response. 19 78
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