Studies on the Polysaccharide-Sulfating System in Sea Urchin Embryos. Ii. Nature of Sulfated Materials Produced in a Cell-Free System*1

Abstract

Sulfated polysaccharides produced by the reaction which transfers sulfate from 3 '-phosphoadenosine 5 '-phosphosulfate to endogenous acceptors in sea urchin embryos were characterized by their precipitability with cetylpyridinium chloride, chromatographic behavior on DEAE-cellulose and Sephadex G-75 columns, and digestibility with chondroitinase ABC and AC.One component of the sulfated polysaccharides was identified as dermatan sulfate (DS). The others were chondroitinase ABC-resistant and separable into large (SPL) and small (SPS) molecular polysaccharides by Sephadex G-75 column chromatography.Sulfate incorporation into SPL was always higher than incorporation into DS or SPS and the activity for sulfate incorporation into DS was lost most rapidly during storage of the particulate fraction at 14°C.Sulfate incorporation into the three components was low at the morula stage and then increased during development to the mesenchyme blastula stage. From the mesenchyme blastula to the pluteus stage, incorporation into SPL increased and that into SPS remained constant. Incorporation into DS was maximal from the blastula to the late gastrula stage and became low at the pluteus stage.GAG has been found in sea urchin embryos (10-15, 17-20, 28, 30) and the possibilities that it may be concerned with cell movement (12, 13, 18,28), protein synthesis (12) and regulation of genomic RNA synthesis (14) have been discussed. Sulfation of GAG has been investigated in various tissues and sulfate transfer from PAPS to various endogenous (4-6, 8, 9, 25-27) and exogenous (1, 7, 21, 29) GAG'S has been demonstrated in cell-free systems. Formation of sulfated polysaccharide by incubation of the particulate fraction of sea urchin embryos with [3sS] PAPS at pH 6.8 has been reported (23). The properties of this polysaccharide-sulfating system in sea urchin embryos were found to be similar to those of GAG-sulfating systems in various tissues (7, 8,29).In the present work, the properties of this polysaccharide-sulfating system were characterized mainly by analyses of the sulfated material produced. The sulfated materials were ** This paper is dedicated to the late Prof. Jean C. Dan.

Sulfated Polysaccharides and Cell Differentiation in the Sea Urchin Embryo

“…Yamagata and Okazaki [1974] and Saotome and Yanagisawa [1979] have shown that der matan sulfate is synthesized in extracts from the embryos of //. pulcherrimus and Anthocidaris crassispina.…”