Collagenases are a category of enzymes that can digest native collagen, the major fibrous component of animal extracellular connective tissue. Collagenases can be produced by some bacterias such as Clostridium and can also be made by the animal body as part of its normal immune response. They normally target the connective tissue in muscle cells and other body organs. They can break peptide bonds within collagen and help create collagen by cleaving pro-collagen, collagen's precursor, once it is secreted by the cell. Bacterial collagenases exhibit broader substrate specificity than animal collagenases and unlike animal collagenases that split collagen in its native triple-helical conformation, bacterial collagenase can degrade both water-insoluble native collagens and water-soluble denatured ones. In addition, bacterial collagenase can attack almost all collagen types, and is able to make multiple cleavages within triple helical regions. Collagenases have a molecular weight ranging from 68 kilodaltons (kDa) to 130 Kda due to differing sizes based on the class to which the isolate belongs. The optimal pH for stability ranges from approximately 6.3 to 7.5 with a theoretical isoelectric point of 5.62. Numerous compounds and molecules have the capability of acting as collagenases activator or inhibitor. For example, Ca2+ and Zn2+ ions are two known activators. On the other hand, ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), cysteine, histidine, dithiothreitol (DTT), 2-mercaptoethanol, o-phenanthroline, Hg2+, Pb2+, Cd2+, Cu2+, and Zn2+ are all known inhibitors of collagenase activity.
1.https://en.wikipedia.org/wiki/Collagenase
2.http://www.wisegeek.com/what-is-collagenase.htm
3.http://www.rockland-inc.com/collagenase.aspx
4.http://www.worthington-biochem.com/cls/default.html
This collagenase has been tested with cell lines to verify the product is not cytotoxic. Collagenase is typically used to digest the connective components in tissue samples to liberate individual cells. The concentration for cartilage dispersal is 1-2 mg/ml, but literature searches should be performed for species specific and/or tissue specific concentrations.
In investigation of the structure and biosynthesis of collagen; in dispersion of cells for tissue culture studies.
Collagenase, Type I, Clostridium histolyticum is used as a powder and solvent injection kit for the treatment of dupuytren's contracture.
A rare proteinase that catalyzes the hydrolysis of collagen. Collagenase activity has been demonstrated
in several venoms of crotalid and viperid snakes and
it occurs in certain species of clostridium.
Collagenase from Clostridium histolyticum comprises of collagenases G and H. Collagenase has N-terminal activator domain and C-terminal peptidase domain. The collagen binding domains (CBD) and polycystic kidney disease-like (PKD-like) domain resides in the peptidase domain. The collagenase module undergoes structural changes upon calcium binding.
Effective release of cells from tissue requires the action of collagenase enzymes and the neutral protease. Collagenase is activated by four gram atom calcium (Ca2+) per mole enzyme. The culture filtrate is thought to contain at least 7 different proteases ranging in molecular weight from 68-130 kDa. The pH optimum is 6.3-8.8. The enzyme is typically used to digest the connective components in tissue samples to liberate individual cells. Ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA)4; β-mercaptoethanol; glutathione, reduced; thioglycolic acid, sodium; and 2,2′-dipyridyl; 8-hydroxyquinoline are known to inhibit the enzyme activity.
Collagenase is purified by using N-ethylmaleimide to activate the enzyme, and wheat germ agglutinin-agarose affinity chromatography [Callaway et al. Biochemistry 25 4757 1986].
