Purification and partial amino acid sequence of osteogenin, a protein initiating bone differentiation.
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.
Luyten
F P
FP
Bone Cell Biology Section, National Institute of Dental Research, Bethesda, Maryland 20892.
Cunningham
N S
NS
Ma
S
S
Muthukumaran
N
N
Hammonds
R G
RG
Nevins
W B
WB
Woods
W I
WI
Reddi
A H
AH
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
D006133:Growth Substances; D006882:Hydroxyapatites; D010446:Peptide Fragments; D011506:Proteins; D017886:Durapatite; D014357:Trypsin