title: Purification and chemical modification of porcine bone morphogenetic protein.
abstract: Implantation of porcine bone morphogenetic protein (pBMP) in the muscle induces differentiation of mesenchymal-type cells and results in endochondral bone formation. pBMP was isolated from porcine demineralized bone matrix and purified by hydroxyapatite chromatography, Sephadex G75 gel filtration, preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), preparative isoelectric focusing (IEF), and chromatofocusing fast protein liquid chromatography (FPLC). Porcine BMP has an MW of 26 K and a range of pI from 4.65 to 4.73 determined by SDS-PAGE and IEF, respectively. Reconstitution with the citrate buffer supernatant fraction enables as little as 50 micrograms of the soluble pBMP fractions to induce osteogenesis in an in vivo assay. Chemical modification studies indicate that the osteoinductive potential of the pBMP molecule depends on tyrosine, carboxyl groups, and disulfide bonds and can be increased by modification of sulfhydryl groups. Modification of arginine and tryptophan has no effect on bioactivity. By pepsin-limited proteolysis, fragments of pBMP with an MW of 6-14 K show definite, although reduced, BMP activity.
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Beijing, People's Republic of China.
authors.forename: L
authors.initials: L
authors.lastname: Ko
authors.forename: G X
authors.initials: GX
authors.lastname: Ma
authors.forename: H L
authors.initials: HL
authors.lastname: Gao
mesh_terms: D000818:Animals; D001681:Biological Assay; D001857:Bone Matrix / Q000187:drug effects / Q000378:metabolism; D019485:Bone Morphogenetic Proteins; D001842:Bone and Bones / Q000187:drug effects / Q000378:metabolism*; D006133:Growth Substances / Q000302:isolation & purification* / Q000494:pharmacology; D008722:Methods; D051379:Mice; D008970:Molecular Weight; D010012:Osteogenesis / Q000187:drug effects; D010434:Pepsin A / Q000494:pharmacology; D011506:Proteins / Q000187:drug effects / Q000302:isolation & purification* / Q000494:pharmacology; D013552:Swine
chemical_list: D019485:Bone Morphogenetic Proteins; D006133:Growth Substances; D011506:Proteins; D010434:Pepsin A