Isolation and characterization of dermatan sulphate proteoglycan fom human uterine cervix

Abstract

Proteoglycans were extracted from human uterine cervix with 4 M-guanidinium chloride in the presence of proteinase inhibitors. They were purified by density-gradient centrifugation in 4 M-guanidinium chloride/CsCl (starting density 1.32 g/ml) followed by DEAE-cellulose and Sepharose chromatography. Only one polydisperse proteoglycan was found. S0[20,w] was 2.1S and the weight-average molecular weight was 73,000 (sedimentation-equilibrium centrifugation) to 110,500 (light-scattering). The core protein was monodisperse, with an apparent molecular weight of 47,000. The proteoglycan contained about 30% protein and probably two or three glycosaminoglycan side chains per molecule. High contents of aspartate, glutamate and leucine were found. The glycan moiety of the proteoglycan was exclusively dermatan sulphate, with a co-polymeric structure with approximately equal quantities of iduronic acid- and glucuronic acid-containing disaccharides.