EDTA-insoluble, calcium-binding proteoglycan in bovine bone
| Autor: | Gayle E. Lester, Bruce Caterson, Y. Hashimoto, Mitsuo Yamauchi |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
Keratan sulfate
Endocrinology Diabetes and Metabolism Blotting Western chemistry.chemical_element Enzyme-Linked Immunosorbent Assay Calcium Bone and Bones chemistry.chemical_compound Endocrinology Aspartic acid medicine Animals Trypsin Orthopedics and Sports Medicine Asparagine Amino Acids Edetic Acid chemistry.chemical_classification Chromatography biology Chemistry Coomassie Brilliant Blue Amino acid Molecular Weight Solubility Biochemistry Proteoglycan Chromatography Gel biology.protein Cattle Electrophoresis Polyacrylamide Gel Proteoglycans medicine.drug |
| Zdroj: | Calcified Tissue International. 56:398-402 |
| ISSN: | 1432-0827 0171-967X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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