The release of collagenase as an inactive proenzyme by bone explants in culture.
1. A latent collagenase, activated only by limited proteolysis, was found in culture media of mouse bone explants. It could be activated by trypsin or, less efficiently, by chymo-trypsin. Skin explants also released latent collagenase. 2. Bone collagenase attacks native collagen at about neutral pH when it is in solution, in reconstituted fibrils or in insoluble fibres, producing two fragments representing 75 and 25% of the molecule. It requires calcium and is inhibited by EDTA, cysteine or serum. 3. Latent collagenase is not activated by trypsin-activated collagenase but by a distinct unidentified thermolabile agent present in a latent trypsin-activatable state in the culture media, or by purified liver lysosomes between pH5.5 and pH7.4. Trypsin activation decreases the molecular weight of latent collagenase from 105000 to 84000 as determined by gel filtration. 5. The latency of collagenase is unlikely to be due to an enzyme-inhibitor complex. Although some culture media contain a collagenase inhibitor, its presence is not constant and its molecular weight (at least 120000) is not compatible with the decrease in molecular weight accompanying activation; also combinations of collagenase with inhibitor are not reactivated by trypsin. Moreover, the latency remains after gel filtration, or treatment by high dilution, exposure to pH values between 2.5 and 10, or high ionic strength, urea or detergent. 6. It is proposed that latent collagenase represents an inactive precursor of the enzyme, a ;procollagenase', and that the extracellular activity of collagenase is controlled by another protease that activates procollagenase by a limited proteolysis of its molecule.
D000818:Animals; D001842:Bone and Bones / Q000201:enzymology*; D002118:Calcium; D002247:Carbon Isotopes; D002850:Chromatography, Gel; D002918:Chymotrypsin; D003094:Collagen / Q000096:biosynthesis; D003470:Culture Media; D046508:Culture Techniques; D003545:Cysteine; D003956:Dialysis; D004492:Edetic Acid; D004586:Electrophoresis; D004622:Embryo, Mammalian / Q000201:enzymology; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism*; D005374:Filtration; D005998:Glycine / Q000378:metabolism; D006168:Guinea Pigs; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000166:cytology; D008247:Lysosomes; D008297:Male; D051379:Mice; D003012:Microbial Collagenase / Q000037:antagonists & inhibitors; D008970:Molecular Weight; D009924:Organ Culture Techniques; D051381:Rats; D012867:Skin / Q000201:enzymology / Q000378:metabolism; D014357:Trypsin
D002247:Carbon Isotopes; D003470:Culture Media; D004792:Enzyme Precursors; D003094:Collagen; D004492:Edetic Acid; D002918:Chymotrypsin; D014357:Trypsin; D003012:Microbial Collagenase; D003545:Cysteine; D002118:Calcium; D005998:Glycine