An active neutral metalloproteinase bound to the insoluble collagen in the mineralized phase matrix of adult rat calvaria.
Two kinds of proteinases were found in the mineralized phase matrix of 24-week-old rat calvaria by means of enzymography using gelatin as a substrate. One proteinase was a neutral thiol 58kD proteinase as shown in a previous paper [2]. The other was a neutral metalloproteinase that had a molecular mass of 56kD and was detected only when calcium (Ca) ions were added to the incubation buffer. It is believed that the 56kD proteinase is bound to the insoluble collagen of the bone matrix, as it is solubilized by 4 M guanidine HC1 solution from the insoluble collagen fraction, when prepared by removing extractable proteins of the mineralized phase matrix. The insoluble collagen fraction could also be solubilized and prepared as gelatin by heating at 65 degrees C for 5 minutes. The gelatin was then incubated at 37 degrees C without further treatment and became degraded without an activation of 4-aminophenylmercuric acetate (APMA). This nonactivated degradation was enhanced by adding Ca ions. These results suggest that the 56kD metalloproteinase bound to the insoluble collagen of bone matrix is in an active form and may participate in the rapid degradation of collagen during bone resorption. As partially purified 56kD metalloproteinase degraded cartilage type proteoglycan, but not type I, IV, and V collagens, it is possibly related to the degradation of proteoglycans before it binds to collagen fibers during bone formation.
Fukae
M
M
Department of Biochemistry, School of Dental Medicine, Tsurumi University, Yokohama, Japan.
Tanabe
T
T
Yamada
M
M
D000818:Animals; D001857:Bone Matrix / Q000201:enzymology*; D001862:Bone Resorption; D002118:Calcium / Q000378:metabolism; D003094:Collagen / Q000302:isolation & purification / Q000378:metabolism*; D005260:Female; D008297:Male; D008666:Metalloendopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D051381:Rats; D012886:Skull / Q000201:enzymology* / Q000378:metabolism
D003094:Collagen; D008666:Metalloendopeptidases; D002118:Calcium