In situ phosphorylation of bone and dentin proteins by the casein kinase II-like enzyme.
Our previous studies suggested the possibility of extracellular phosphorylation of matrix phosphoproteins into more phosphorylated forms by mature odontoblasts and osteocytes (Mikuni-Takagi et al., 1995; Satoyoshi et al., 1995). To elucidate such phosphorylation of bone and dentin proteins, we developed a histochemical method using frozen sections to determine the sites of enzymatic processing by the casein kinase II-like enzyme. It was observed that proteins in bone, dentin, and predentin are phosphorylated by the endogenous enzyme when the tissue slices were incubated with [gamma-32P] GTP, suggesting that there are both substrates and the enzyme in these matrices. In vivo, phosphate donors, ATP and GTP, may be supplied through dentinal canals and osteocyte canaliculi. Immunohistochemical analysis of frozen sections showed that the extremely intense staining of phosphoserine residues by anti-phosphoserine antibodies appeared in dentin only after demineralization of the tissue samples. It implies that these phosphoserine residues become bound to mineral as soon as the phosphorylation is completed, thereby being inaccessible to the antibodies without demineralization. The data support our notion that the extracellular phosphorylation of dentin/bone proteins, regulated by the developmental stages of bone and dentin cells, occurs prior to matrix mineralization.
Suzuki
Y
Y
Department of Oral Biochemistry, Kanagawa Dental College, Yokosuka, Japan.
Yamaguchi
A
A
Ikeda
T
T
Kawase
T
T
Saito
S
S
Mikuni-Takagaki
Y
Y
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001857:Bone Matrix / Q000378:metabolism*; D002113:Calcification, Physiologic / Q000502:physiology; D047390:Casein Kinase II; D004268:DNA-Binding Proteins / Q000378:metabolism*; D003804:Dentin / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism; D007150:Immunohistochemistry; D008903:Minerals / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
D004268:DNA-Binding Proteins; D008903:Minerals; D010750:Phosphoproteins; C017630:phosphophoryn; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D000469:Alkaline Phosphatase