Features of Collagen Matrix Reconstructed with Proteodermatan sulfate from Pigskin Insoluble Collagen

Abstract

Pigskin insoluble collagen was step-wise extracted with 0.5 M and 4M guanidine hydrochloride (GuHCl) solution. Two proteoglycan preparations were obtained by DEAE-Toyopearl chromatography of the extracts, PG-I from 0.5 M GuHCl extract and PG-II from 4M GuHCl extract. The proteoglycan preparations both contained at least two components having different mobilities on SDS-PAGE (M_r, about 60 × 10³ and 90 × 10³), and were identified as proteodermatan sulfate (PDS) by two-dimensional electrophoresis on a cellulose acetate membrane. The two proteodermatan sulfates (PG-I and PG-II) accelerated the rate of the reconstruction of collagen matrix from neutral collagen solution, while the accelerating effect of PG-I was greater than that of PG-II. Electron microscopic observation showed that collagen matrices reconstructed with the PDSs consisted of thick fibrils with the banding pattern of native type and thin fibrils with an indistinct banding pattern. Collagen matrix reconstructed with addition of PDSs showed lower denaturation temperature than the control, as assessed by differential scanning calorimetry. These results suggest that collagen tissue-bound PDSs have a regulative function in the reconstruction of collagen matrix and its thermal stability.