EDTA-insoluble, calcium-binding proteoglycan in bovine bone.
A calcium ion precipitable, trypsin-generated proteoglycan fragment has been isolated from the demineralized, EDTA-insoluble matrices of bone. The demineralized matrix was completely digested with trypsin, increasing concentrations of CaCl2 were added to the supernatant, and the resulting precipitates were analyzed. The amount of precipitate gradually increased with higher concentrations of calcium and was reversibly solubilized by EDTA. After molecular sieve and anion exchange chromatography, a proteoglycan-containing peak was obtained. Immunochemical analysis showed that this peak contained chondroitin 4-sulfate and possibly keratan sulfate. Amino acid analysis showed that this proteoglycan contained high amounts of aspartic acid/asparagine (Asx), serine (Ser), glutamic acid/glutamine (Glx), proline (Pro), and glycine (Gly); however, it contained little leucine (Leu) which suggests that it is not a member of the leucine-rich small proteoglycan family. In addition, significant amounts of phosphoserine (P-Ser) and hydroxyproline (Hyp) were identified in hydrolysates of this fraction. A single band (M(r) 59 kDa) was obtained on SDS-PAGE that stained with Stains-all but not with Coomassie Brilliant Blue R-250. If bone powder was trypsinized prior to demineralization, this proteoglycan-containing fraction was not liberated. Collectively, these results indicate that a proteoglycan occurs in the demineralized matrix that is precipitated with CaCl2 and is closely associated with both mineral and collagen matrices. Such a molecule might facilitate the structural network for the induction of mineralization in bone.
Hashimoto
Y
Y
Department of Biochemistry, School of Dentistry, Aichi-gakuin University, Nagoya, Japan.
Lester
G E
GE
Caterson
B
B
Yamauchi
M
M
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D015153:Blotting, Western; D001842:Bone and Bones / Q000737:chemistry*; D002118:Calcium / Q000378:metabolism*; D002417:Cattle; D002850:Chromatography, Gel; D004492:Edetic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D008970:Molecular Weight; D011509:Proteoglycans / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility; D014357:Trypsin
D000596:Amino Acids; D011509:Proteoglycans; D004492:Edetic Acid; D014357:Trypsin; D002118:Calcium