Purification of Water-Soluble Bone-Inductive Protein from Bovine Demineralized Bone Matrix
| Autor: | Jun Kawada, Naoyuki Matsumoto, Kikuji Yamashita, Yoshiyuki Yoshimura, Yasuyuki Horisaka, Tomomichi Takagi, Mikio Nishida, Atsuko Hirano, Yasuo Okamoto |
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| Rok vydání: | 1993 |
| Předmět: |
Male
Bone Matrix Ultrafiltration Pharmaceutical Science chemistry.chemical_element Calcium Bone and Bones Chromatography Affinity Gel permeation chromatography Affinity chromatography Protein purification medicine Animals Rats Wistar Pharmacology Gel electrophoresis Bone Development Chromatography Chemistry Demineralized bone matrix Proteins Osteoblast General Medicine Alkaline Phosphatase Rats medicine.anatomical_structure Biochemistry Bone Morphogenetic Proteins Chromatography Gel Alkaline phosphatase Cattle Electrophoresis Polyacrylamide Gel |
| Zdroj: | Biological and Pharmaceutical Bulletin. 16:444-447 |
| ISSN: | 1347-5215 0918-6158 |
| Popis: | The water-soluble fraction containing bone-inductive activity was purified from guanidine-hydrochloride extracts of bovine demineralized bone. The purification steps include ultrafiltration, dialysis, affinity chromatography on heparin-Sepharose and gel chromatography on Sephacryl S-200. Combination of these steps was proven to be an effective and rapid method for the purification of this protein. Subcutaneous implantation of the water-soluble protein with type I collagen was carried out in the thorax of rats. When alkaline phosphatase activity and calcium content in implants were used as indices for purification, the water-soluble bone-inductive protein was purified600-fold according to the enzyme activity and 64-fold according to the calcium content. A morphological examination revealed that many chondrocyte and osteoblast cells were seen in the location of the implanted material. Sodium dodecyl sulfate/gel electrophoresis of the protein produced in this way under non-reducing conditions revealed four protein bands of 18, 16, 14 and 11 kDa. None of the separated bands had any biological activity. This result suggests that the water-soluble bone-inductive activity depends on an associated form of various proteins in the range of 18 to 11 kDa. |
| Databáze: | OpenAIRE |
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