Characterization of fibromodulin isolated from bovine periodontal ligament.
Although several proteoglycans (PGs) have been reported in bovine periodontal ligament (PDL), the composition of PGs in PDL has been poorly characterized. In the present study, we isolated and characterized keratan sulfate-substituted PG (fibromodulin) in bovine PDL. Fibromodulin was purified from 4 M guanidine hydrochloride (GdmCl) extracts of bovine PDL tissues using DEAE Sephacel ion-exchange chromatography and preparative electrophoresis. Fibromodulin appeared as a single polydisperse band with an apparent molecular weight (MW) of 80,000 (80 kDa) on SDS-PAGE. Digestion of fibromodulin with keratanase or neuraminidase reduced the apparent molecular size, and N-glycanase treatment produced core protein bands of around 40 kDa. Fibromodulin reacted with keratan sulfate monoclonal antibody (5D4) and fibromodulin polyclonal antibodies (alpha-FM). The keratanase-digested fibromodulin reacted with alpha-FM, but not with 5D4. These data suggest that fibromodulin is one of the small PGs in the PDL-matrix and may fulfill construction and maintenance functions in this tissue.
Watanabe
T
T
Department of Orthodontics, Kanagawa Dental College, Yokosuka, Japan.
Kubota
T
T
D000581:Amidohydrolases / Q000737:chemistry; D000818:Animals; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D002417:Cattle; D002847:Chromatography, Agarose; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D004586:Electrophoresis; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D016326:Extracellular Matrix Proteins; D000071235:Fibromodulin; D006026:Glycoside Hydrolases; D019791:Guanidine / Q000737:chemistry; D015151:Immunoblotting; D007632:Keratan Sulfate / Q000032:analysis / Q000276:immunology; D015394:Molecular Structure; D008970:Molecular Weight; D009439:Neuraminidase / Q000737:chemistry; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D010513:Periodontal Ligament / Q000737:chemistry*; D011509:Proteoglycans / Q000032:analysis; D012967:Sodium Dodecyl Sulfate; D001616:beta-Galactosidase / Q000737:chemistry
D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D016326:Extracellular Matrix Proteins; D011509:Proteoglycans; D000071235:Fibromodulin; D012967:Sodium Dodecyl Sulfate; D007632:Keratan Sulfate; D006026:Glycoside Hydrolases; C026093:keratan-sulfate endo-1,4-beta-galactosidase; D009439:Neuraminidase; D001616:beta-Galactosidase; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D019791:Guanidine