title: Purification and partial amino acid sequence of osteogenin, a protein initiating bone differentiation.
abstract: Osteogenin was purified from bovine bone matrix and its activity monitored by an in vivo bone induction assay. The purification method utilized extraction of the bone-inducing activity with 6 M urea, followed by chromatography on heparin-Sepharose, hydroxyapatite, and Sephacryl S-200. Active fractions were further purified by preparative sodium dodecyl sulfate gel electrophoresis without reduction. Osteogenin activity was localized in a zone between 30 and 40 kDa. The amino acid sequences of a number of tryptic peptides of the gel-eluted material were determined. Reduction and alkylation of purified osteogenin in 7 M guanidine hydrochloride resulted in the total loss of biological activity. Sodium dodecyl sulfate gel electrophoresis under reducing conditions revealed a broad band with an apparent molecular mass of 22 kDa.
authors.affiliation: Bone Cell Biology Section, National Institute of Dental Research, Bethesda, Maryland 20892.
authors.forename: F P
authors.initials: FP
authors.lastname: Luyten
authors.forename: N S
authors.initials: NS
authors.lastname: Cunningham
authors.forename: S
authors.initials: S
authors.lastname: Ma
authors.forename: N
authors.initials: N
authors.lastname: Muthukumaran
authors.forename: R G
authors.initials: RG
authors.lastname: Hammonds
authors.forename: W B
authors.initials: WB
authors.lastname: Nevins
authors.forename: W I
authors.initials: WI
authors.lastname: Woods
authors.forename: A H
authors.initials: AH
authors.lastname: Reddi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001842:Bone and Bones / Q000166:cytology*; D002417:Cattle; D002454:Cell Differentiation; D002845:Chromatography; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D017886:Durapatite; D006133:Growth Substances / Q000235:genetics / Q000302:isolation & purification*; D006882:Hydroxyapatites; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010012:Osteogenesis; D010446:Peptide Fragments / Q000302:isolation & purification; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification*; D014357:Trypsin
chemical_list: D006133:Growth Substances; D006882:Hydroxyapatites; D010446:Peptide Fragments; D011506:Proteins; D017886:Durapatite; D014357:Trypsin