Isolation of proteoglycan (versican) aggregate from rat dental pulp.
Versican is a large interstitial proteoglycan that is believed to be able to bind hyaluronan to form large aggregate structures, but no study has isolated native versican aggregates from any tissue. In this study, ternary aggregate structures consisting of versican, hyaluronan, and link protein were isolated from rat dental pulp by associative extractions followed by caesium sulphate rate zonal sedimentation centrifugation. Fractions from the centrifugation were analysed by dot blot and Western blot using monoclonal antibodies and hyaluronan-binding protein. About 60% of the hexuronic acid was extracted by associative extractions. Positive reactions for versican, hyaluronan and link protein were clearly detected in the bottom fractions from the centrifugation, but were barely detectable in the top fractions. These results suggest that the majority of the versican, hyaluronan, and link protein forms ternary aggregate structures in the rat dental pulp.
Shibata
S
S
Department of Maxillofacial Anatomy, Graduate School, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, Japan. S.Shibata.oan1@dent.tmd.ac.jp
Yoneda
S
S
Yanagishita
M
M
Yamashita
Y
Y
D000818:Animals; D015153:Blotting, Western / Q000379:methods; D011508:Chondroitin Sulfate Proteoglycans / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D003782:Dental Pulp / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D016326:Extracellular Matrix Proteins; D006820:Hyaluronic Acid / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D015151:Immunoblotting / Q000379:methods; D037181:Lectins, C-Type; D011506:Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D011509:Proteoglycans / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D053675:Versicans; D000099249:Proteoglycan Link Protein
D011508:Chondroitin Sulfate Proteoglycans; D016326:Extracellular Matrix Proteins; D006820:Hyaluronic Acid; D037181:Lectins, C-Type; D011506:Proteins; D011509:Proteoglycans; D053675:Versicans; D000099249:Proteoglycan Link Protein; C508324:Vcan protein, rat