Isolation and partial characterization of a growth factor from human cementum.
Cementum is the mineralized interface through which collagen fibers of periodontal connective tissues are anchored onto the tooth surface. We have isolated and partially characterized a mitogenic factor from human cementum which has properties different from other growth factors. Cementum was harvested from healthy human teeth, extracted in 1.0 M CH3COOH and mitogenic activities were fractionated by heparin-affinity chromatography. Proteins eluted by 0.4-0.6 M NaCl, which contained most of the cementum mitogenic activity, were precipitated by trichloroacetic acid and resolved by HPLC through ion-exchange and reverse-phase columns. NaDodSO4-polyacrylamide gel electrophoresis revealed that the purified preparation contained a M(r) 23,000 protein and this protein was associated with mitogenic activity. The purified cementum-derived growth factor (CGF) was active alone, but at suboptimal concentrations its activity was potentiated by small quantities of plasma-derived serum and epidermal growth factor (EGF). The activity was resistant to heat, but it was destroyed by trypsin digestion. Reduction and alkylation destroyed the mitogenic activity, however electrophoretic mobility was not affected. Binding of EGF to fibroblast membranes was not affected by the CGF and assays to detect platelet-derived growth factor were negative. These characteristics indicated that CGF is a distinct molecular species. Our data show that cementum contains several mitogenic factors and that CGF is the major cementum mitogen.
Yonemura
K
K
Department of Pathology, School of Medicine, University of Washington, Seattle.
Narayanan
A S
AS
Miki
Y
Y
Page
R C
RC
Okada
H
H
D002455:Cell Division; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003739:Dental Cementum / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D006133:Growth Substances / Q000302:isolation & purification* / Q000494:pharmacology; D006801:Humans; D008934:Mitogens / Q000302:isolation & purification* / Q000494:pharmacology; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology
D006133:Growth Substances; D008934:Mitogens; D010982:Platelet-Derived Growth Factor; D004815:Epidermal Growth Factor