Matrix metalloproteinase-2 in dentin matrix mineralization.
In the serum-free culture medium of bovine odontoblasts we detected active gelatinolytic metalloproteinases, matrix metalloproteinase (MMP)-2 and MMP-9 (gelatinases A and B). The activity of MMP-2, in particular, appeared suddenly around day 21 in the culture, coinciding with the development of odontoblastic cell processes and the loss of alkaline phosphatase. Reverse transcriptase-polymerase chain reaction analysis of these odontoblasts demonstrated that messages of MMP-2 but not MMP-9 increased significantly between day 15 and day 21. The in vitro observation indicates that medium conditioned by these odontoblasts and containing significant amounts of MMP-2 degrades not only the collagenous substrates but also purified dentin phosphophoryn as well. We have also observed that dephosphorylated dentin phosphoprotein becomes a better substrate for casein kinase II after limited proteolysis with MMP-2. These results support our working hypothesis that MMP-2-mediated proteolytic processing is an important step in accelerating the process of dentin matrix maturation, which includes phosphorylation and subsequent mineralization. As has been suggested previously, extracellular phosphorylation of matrix proteins is an important step in biomineralization both in bone and in dentin (Mikuni-Takagaki et al., J Bone Miner Res 1995;10:231-42; Zhu et al., Biochem J 1997; 323:637-43). Our present histochemical analysis in MMP-2 knockout mice confirms the concept with the delayed formation of mineralized tissues, dentin, and bone.
Satoyoshi
M
M
Department of Operative Dentistry and Endodontics, Kanagawa Dental College, Yokosuka, Japan.
Kawata
A
A
Koizumi
T
T
Inoue
K
K
Itohara
S
S
Teranaka
T
T
Mikuni-Takagaki
Y
Y
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D047388:Casein Kinases; D002417:Cattle; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D003804:Dentin / Q000201:enzymology* / Q000502:physiology; D007527:Isoenzymes / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000032:analysis* / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000032:analysis / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D009804:Odontoblasts / Q000166:cytology / Q000201:enzymology; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014074:Tooth Calcification / Q000502:physiology*
D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D007527:Isoenzymes; D010750:Phosphoproteins; C017630:phosphophoryn; D003094:Collagen; D011494:Protein Kinases; D047388:Casein Kinases; D000469:Alkaline Phosphatase; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9