Isolation of proteoglycan (versican) aggregate from rat dental pulp

Versican is a large interstitial proteoglycan that is believed to be able t o bind hyaluronan to form large aggregate structures, but no study has isol ated native versican aggregates from any tissue. In this study, ternary agg regate structures consisting of versican, hyaluronan, and link protein were isolated from rat dental pulp by associative extractions followed by caesi um sulphate rate zonal sedimentation centrifugation. Fractions from the cen trifugation were analysed by dot blot and Western blot using monoclonal ant ibodies and hyaluronan-binding protein. Abc ut 60% of the hexuronic acid wa s extracted by associative extractions. Positive reactions for versican, hy aluronan and link protein were clearly detected in the bottom fractions fro m the centrifugation, but were barely detectable in the top fractions. Thes e results suggest that the majority of the versican, hyaluronan, and link p rotein forms ternary aggregate structures in the rat dental pulp. (C) 2000 Elsevier Science Ltd. All rights reserved.