title: Isolation and partial characterization of heparan sulphate proteoglycans from human hepatic amyloid.
abstract: Proteoglycans were isolated from human amyloidotic liver by extraction with guanidine, followed by trichloroacetic acid precipitation, DEAE-Sephacel ion-exchange chromatography, and Sepharose CL-6B gel chromatography. A significant portion of the material was found to be free chondroitin/dermatan sulphate chains (30%), whereas the predominant part was heparan sulphate proteoglycan (HSPG) (70%). The approx. molecular mass of the HSPG was 200 kDa, as measured by gel electrophoresis and gel chromatography. The molecular mass of the core protein was shown to be 60 kDa by SDS/PAGE following de-aminative cleavage of the heparan sulphate chains. The heparan sulphate chains were liberated from the core protein by alkali treatment and found to have a molecular mass of approx. 35 kDa by Sepharose CL-6B gel chromatography. The core protein was shown, by immunoblotting, to react with a monoclonal antibody against bovine basement membrane HSPG. The presence of HSPG in amyloid deposits was further confirmed by immunohistochemistry on tissue sections from amyloidotic liver using the same antibody.
authors.affiliation: Department of Rheumatology, University of Tromsø, Norway.
authors.forename: J H
authors.initials: JH
authors.lastname: Magnus
authors.forename: T
authors.initials: T
authors.lastname: Stenstad
authors.forename: G
authors.initials: G
authors.lastname: Husby
authors.forename: S O
authors.initials: SO
authors.lastname: Kolset
mesh_terms: D000682:Amyloid / Q000737:chemistry*; D000686:Amyloidosis / Q000378:metabolism*; D002648:Child; D002675:Child, Preschool; D002845:Chromatography; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000737:chemistry; D008107:Liver Diseases / Q000378:metabolism*; D008297:Male; D008970:Molecular Weight; D011134:Polysaccharides / Q000302:isolation & purification; D011509:Proteoglycans / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D000682:Amyloid; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D011134:Polysaccharides; D011509:Proteoglycans