Tadpole Collagenase. Preparation and Purification<sup>*</sup>

Nagai, Yutaka; Lapière, Ch. M.; Gross, Jerome

doi:10.1021/bi00874a007

Cited by 384 publications

(93 citation statements)

References 14 publications

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“…Since tissue collagenases are digestive enzymes, they are commonly isolated from the digestive tracts of various fish and invertebrates including: tadpole tailfin (Gross and Nagai, 1965;Nagai et al, 1966), rabbit skin (Fullmer and Gibson, 1966), rat uterus (Jeffrey and Gross, 1967), rheumatoid synovial tissue (Evanson et al, 1968), mouse bones (Sakamoto et al, 1972), crabs (Eizen and Jeffrey, 1969;Grant et al, 1983;Klimova et al, 1990;Sellos and Van Wormhoudt, 1992;Gerasimova and Kupina, 1996;Zefirova et al, 1996), fresh water prawns (Baranowski et al, 1984), crayfish (Garcia-Carreno et al, 1994), Atlantic cod (Kristjánsson, et al, 1995), tropical shrimp (Penaeus vannamei) Van Wormhoudt et al, 1992) and catfish (Parasilurus asotus) (Klimova et al,1990;Sellos and Van Wormhoudt, 1992).…”

Section: Collagenase and Collagenolytic Enzymesmentioning

confidence: 99%

Extraction and Purification of Collagenase Enzymes: A Critical Review

Daboor¹,

Budge²,

Ghaly

et al. 2010

American Journal of Biochemistry and Biotechnology

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Problem statement: Enzymes have vital roles in several industrial processes (foods, cosmetics, nutraceuticals and pharmaceuticals) due to their highly selective nature and high activity at very low concentrations. Recent efforts to identify new sources of useful enzymes have been concentrated on the marine environment because of the potential to make use of processing wastes. About 35-50% of the mass of the fish caught is a waste that is disposed off at sea or in landfills. The extraction of enzymes from fish processing waste can reduce environment problems and improve the economics of the fish industry. Collagenases are a group of enzymes that can be extracted from fish waste. Approach: Comprehensive reviews of the literature on the extraction, purification, characterization and use of collagenases was carried out. Results: Collagenases have different molecular weights based on their types and sources. They have the ability to break down the peptide bonds in collagen at physiological pH. They are classified into two types: serine and metallocollagenase. Collagenolytic activities have been shown at a wide range of temperatures (20-40°C) and pH (6-8). Many activators can be used to achive collagenase activity including 4-Aminophenylmercuric Acetate (APMA), trypsin, potassium or sodium thiocyanate, iodoacetamide and potassium iodide. Dithiothreitol (DTT), mercaptoethanol, ethylendiaminetetracetic acid, ophenanthroline and cysteine inactivate the enzyme. Collagenases enzymes can be extracted with a variety of techniques using different buffering systems (tris-HCl, sodium bicarbonate, calcium chloride and cacodylate). All techniques involve the use of ammonium sulphate fractionation and centrifugation to precipitate the enzyme. Collagenases are normally purified using chromatographic techniques such as gel-filtration, ion-exchange and affinity column chromatography. Collagenase can be assayed with a number of methods, including: colorimetric absorbance, viscometry, radioactivity and fluorescence spectroscopy. Collagenases are partly responsible for toughness in red meats and are used as tenderizers in food industry, have application in the fur and hide tanning to ensure uniform dying of leather, used in medicine to treat burns and ulcers, eliminate scar tissues, transplantation of organs. Conclusion: Understanding of the nature of the enzymes and identifying the most suitable resources and the methods for their extraction and purification will have significant impact on the fish processing, food and medical industries.

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Section: Collagenase and Collagenolytic Enzymesmentioning

confidence: 99%

Extraction and Purification of Collagenase Enzymes: A Critical Review

Daboor¹,

Budge²,

Ghaly

et al. 2010

American Journal of Biochemistry and Biotechnology

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“…Collagenolytic activity was assayed by measuring the release of soluble radioactive products from a pellet of thermally reconstituted ['4C]glycine-labelled collagen fibrils by a modification of the method of Nagai et al [16].…”

Section: Collagenase Assaymentioning

confidence: 99%

Action of Rheumatoid Synovial Collagenase on Cartilage Caollagen

Woolley

Glanville

Evanson

et al. 1975

European Journal of Biochemistry

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The action of purified rheumatoid synovial collagenase on purified cartilage collagen, U~( I I )~, in solution at 25 "C has been characterised. The enzyme attacked cartilage collagen in solution producing a 5 8 % reduction in specific viscosity and resulting in the appearance of two reaction products which represented approximately three-quarter and one-quarter fragments of the intact molecule as shown by disc electrophoresis in polyacrylamide gels containing sodium dodecyl sulphate. The a-chain fragments which comprised each of these components corresponded to molecular weights of approximately 74000 and 21 000.Electron microscopy of segment-long-spacing crystallites of the reaction products revealed three-quarter (TCA) and one-quarter (TCB) length fragments, and permitted accurate localization of the cleavage locus between bands41 and 42 (1-41). This cleavage site and the formation of TCA and TCB reaction products are very similar to those found for type-I collagen substrates.Cartilage collagen in solution was found to be more resistant to collagenase attack than tendon collagen, the rate of cartilage collagen degradation being six times slower than that for tendon collagen, as judged by viscometry. The mid-point melting temperatures (T,) for lathyritic cartilage and tendon collagen were 40.5 and 41.5 "C, and for the collagenase-produced reaction products 38.5 and 37.5 "C, respectively. The significance of these findings is discussed in relation to the structure of type I and I1 collagens.At 25 "C rheumatoid synovial collagenase, like other mammalian collagenases, has been shown to attack the collagen molecule in its native triple-helical conformation at a specific locus [l-31 yielding reaction products, commonly known as TCA and TCB, which represent three-quarter and one-quarter length fragments of the collagen molecule respectively. These products have been shown to be more susceptible than the native molecules to thermal denaturation [l]. Such findings were obtained using solutions of mammalian, type I, skin collagen having the molecular composition al(I)2a2, whereas collagen extracted from cartilage containing three identical a chains, and having a molecular composition al(II), [4-61, has been reported to be resistant to collagenases derived from gingival and polymorphonuclear leucocytes [7].Since earlier studies had shown that crude preparations of synovial collagenase were able to digest whole slices of articular cartilage [8,9] and more Enzyme. Collagenase (EC 3.4.24.3); trypsin (EC 3.4.21.4).Eur. J. Biochem. 50 (1975) recently, cartilage collagen has been shown to be attacked by rabbit corneal collagenase [lo] and human skin collagenase [l 13, we have compared the action of purified synovial collagenase on purified cartilage (type 11) and tendon (type I) collagens from lathyritic chicks. MATERIALS AND METHODS Preparation and Purification of Rheumatoid Synovial CollagenaseRheumatoid synovial tissue obtained at synovectomies was cut into fragments approximately 4 mm in greatest dimension and explante...

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“…This led to the discovery of amphibian collagenase-I, which founded the protease family now commonly known as matrix metalloproteinases (MMPs) (Brinckerhoff and Matrisian, 2002;Nagai et al, 1966). Since the discovery that proteases are linked to tissue remodeling, the interest in this connection has been constantly growing.…”

Section: Introductionmentioning

confidence: 99%

Interdependency of cell adhesion, force generation and extracellular proteolysis in matrix remodeling

Kirmse

Otto

Ludwig

2011

Journal of Cell Science

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SummaryIt is becoming increasingly evident that the micromechanics of cells and their environment determine cell fate and function as much as soluble molecular factors do. We hypothesized that extracellular matrix proteolysis by membrane type 1 matrix metalloproteinase (MT1-MMP) depends on adhesion, force generation and rigidity sensing of the cell. Melanoma cells (MV3 clone) stably transfected with MT1-MMP, or the empty vector as a control, served as the model system. 21 integrins (cell adhesion), actin and myosin II (force generation and rigidity sensing) were blocked by their corresponding inhibitors (21 integrin antibodies, Cytochalasin D, blebbistatin). A novel, anisotropic matrix array of parallel, fluorescently labeled collagen-I fibrils was used. Cleavage and bundling of the collagen-I fibrils, and spreading and durotaxis of the cells on this matrix array could be readily discerned and quantified by a combined set-up for fluorescence and atomic force microscopy. In short, expression of the protease resulted in the generation of structural matrix defects, clearly indicated by gaps in the collagen lattice and loose fiber bundles. This key feature of matrix remodeling depended essentially on the functionality of 21 integrin, the actin filament network and myosin II motor activity. Interference with any of these negatively impacted matrix cleavage and three-dimensional matrix entanglement of cells.

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Tadpole Collagenase. Preparation and Purification^*

Cited by 384 publications

References 14 publications

Extraction and Purification of Collagenase Enzymes: A Critical Review

Extraction and Purification of Collagenase Enzymes: A Critical Review

Action of Rheumatoid Synovial Collagenase on Cartilage Caollagen

Interdependency of cell adhesion, force generation and extracellular proteolysis in matrix remodeling

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Tadpole Collagenase. Preparation and Purification*

Cited by 384 publications

References 14 publications

Extraction and Purification of Collagenase Enzymes: A Critical Review

Extraction and Purification of Collagenase Enzymes: A Critical Review

Action of Rheumatoid Synovial Collagenase on Cartilage Caollagen

Interdependency of cell adhesion, force generation and extracellular proteolysis in matrix remodeling

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Product

Resources

About

Tadpole Collagenase. Preparation and Purification^*