title: Immunoelectron microscopy of proteodermatan sulfate in human mid-dermis.
abstract: Proteodermatan sulfate was isolated from the skin of human, female breast in 6-M urea and proteolytic inhibitors at 70 degrees C and purified on Sephacryl S-200. It was composed of 55% protein and 45% dermatan sulfate, displayed one protein and carbohydrate-stainable band on agarose-polyacrylamide gels, yielded dermatan sulfate after digestion by papain, and its calculated E0.1% 1 cm, 280 nm was 16.2. Its mucopolysaccharide portion was digested by chondroitinase ABC but not by chondroitinase AC. This proteoglycan was used to immunize rabbits. Double diffusion of antiserum against the antigen or its core protein resulted in one precipitation band. Antiserum did not cross-react with bovine collagen type I, human fibronectin, dermatan sulfate, hyaluronic acid, heparin, heparan sulfate or the chondroitin sulfates by double diffusion. The antiserum titer determined by radioimmunoassay was 1:16,000. This assay was not affected by a 40-fold excess of dermatan sulfate. Purified IgG molecules were apparently associated with collagen in human breast mid-dermis as demonstrated by indirect immunoelectron microscopy with ferritin-labeled goat antirabbit IgG. The results indicate that rabbit anti-human, anti-proteodermatan sulfate IgG is highly specific for the core protein of dermatan sulfate and confirm the hypothesis that in vivo proteodermatan sulfate is closely associated with collagen.
authors.forename: M O
authors.initials: MO
authors.lastname: Longas
authors.forename: R
authors.initials: R
authors.lastname: Fleischmajer
mesh_terms: D002807:Chondroitin / Q000031:analogs & derivatives*; D002850:Chromatography, Gel; D003871:Dermatan Sulfate / Q000031:analogs & derivatives* / Q000032:analysis; D004588:Electrophoresis, Cellulose Acetate; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D005779:Immunodiffusion; D007158:Immunologic Techniques; D008854:Microscopy, Electron; D011509:Proteoglycans / Q000032:analysis*; D012867:Skin / Q000032:analysis*
chemical_list: D011509:Proteoglycans; C028774:proteodermatan sulfate; D003871:Dermatan Sulfate; D002807:Chondroitin