Isolation and characterization of dermatan sulphate and heparan sulphate proteoglycans from fibroblast culture

Publisher Website
Google Scholar
Abstract
35SO42-- and [3H]leucine-labeled proteoglycans were isolated from the medium and cell layer of human skin fibroblast cultures. Measures were taken to avoid proteolytic modifications during isolation by adding guanidinium chloride and proteolysis inhibitors immediately after harvest. The proteoglycans were purified and fractionated by density gradient centrifugation, followed by gel and ion-exchange chromatography. This procedure permitted the isolation of 2 major proteoglycan fractions from the medium, one large, containing glucuronic acid-rich dermatan sulfate chains, and one small, containing iduronic acid-rich ones. The protein core of the latter proteoglycan had an apparent MW of 47,000 as determined by polyacrylamide-gel electrophoresis; protein core of the former was considerably larger. The major dermatan sulfate proteoglycan of the cell layer was similar to the large proteoglycan of the medium. Only small amounts of the iduronic acid-rich dermatan sulfate proteoglycan could be isolated from the cell layer. Most of the iduronic acid-rich glycans appeared as free chains. The heparan sulfate proteoglycans in the cell culture were largely confined to the cell layer. This proteoglycan was of rather low buoyant density and seemed to contain a high proportion of protein. The major part of the heparan sulfate proteoglycan from the medium had a higher buoyant density and contained a smaller amount of protein.

This publication has 13 references indexed in Scilit: