Synthesis of collagenase and collagenase inhibitors by osteoblast-like cells in culture.
A rat osteosarcoma cell clone (ROS 17/2), and osteoblast-enriched populations from rat calvaria cultured in the presence of concanavalin A, have been shown to produce latent collagenase and collagenase inhibitors. The enzymes and inhibitor activities from the ROS 17/2 cells were concentrated by ammonium sulphate precipitation and separated by gel filtration on AcA 54 resin. The size of the latent collagenase (Mr approximately equal to 58000) was reduced on conversion to active enzyme (Mr approximately equal to 48000) by p-aminophenylmercuric acetate. Latent and active forms of gelatinase activity, similar in size to the corresponding forms of collagenase, were also resolved. The collagenase inhibitor activity, which was sensitive to organomercurials, was recovered in two peaks (Mr approximately equal to 68000 and 30000). The active collagenase cleaved interstitial collagens (type I = III greater than II) producing typical 3/4 and 1/4 fragments. This activity was inhibited by the metal ion chelators ethylenediaminetetraacetic acid and o-phenanthroline. Additional specific cleavages of native collagen were also observed which, from the susceptibility of this activity to phenylmethylsulphonyl fluoride, leupeptin and antipain, suggested the presence of a second collagenolytic enzyme. This synthesis of collagenolytic enzymes by these osteoblast-like cells suggests that individual osteoblasts, like fibroblasts, are capable of both synthesizing and degrading their respective organic matrices in vivo.
D000645:Ammonium Sulfate; D000818:Animals; D002460:Cell Line; D055598:Chemical Phenomena; D002621:Chemistry; D002999:Clone Cells; D005333:Fetus; D003012:Microbial Collagenase / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D010006:Osteoblasts / Q000201:enzymology / Q000378:metabolism*; D012516:Osteosarcoma; D010779:Photofluorography; D051381:Rats; D012513:Sarcoma, Experimental; D012886:Skull
D003012:Microbial Collagenase; D000645:Ammonium Sulfate