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Purification of Amyloid‐Associated Heparan Sulphate Proteoglycans and Galactosaminoglycan Free Chains from Human Tissues
Author(s) -
STENSTAD T.,
MAGNUS J. H.,
HUSBY G.,
KOLSET S. O.
Publication year - 1993
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1993.tb01760.x
Subject(s) - perlecan , proteoglycan , basement membrane , chemistry , microbiology and biotechnology , immunohistochemistry , antibody , monoclonal antibody , amyloid (mycology) , kidney , spleen , amyloidosis , blot , staining , glomerular basement membrane , biochemistry , extracellular matrix , pathology , biology , immunology , medicine , glomerulonephritis , endocrinology , inorganic chemistry , gene
Basement membrane‐associated heparan sulphate proteoglycans have been demonstrated immunohistochemically in organs from patients afflicted with various types of amyloidosis. In a recent report, we were able to isolate and partly characterize a basement membrane‐associated heparan sulphate proteoglycan from human hepatic amyloid. In the present study proteoglycans were extracted with guanidine from human amyloid‐laden kidney, spleen and lymph nodes. All tissues extracted with guanidine contained both heparan sulphate proteoglycan (HSPG) and galactosaminoglycan (CS/DS) free chains. Tissue staining using a monoclonal antibody against basement membrane HSPG revealed the presence of HSPG in amyloid deposits in kidney and spleen. Furthermore, following SDS‐PAGE of HSPG from kidney after deaminative cleavage of the HS chains, a 15‐kDa and 80‐kDa protein appeared, probably representing the core protein(s). In lymph node HSPG, three core proteins of 65, 30 and 25 kDa could be demonstrated on SDS‐PAGE, the first reacting with the anti‐basement membrane HSPG antibody when subjected to Western blotting subsequent to SDS‐PAGE. By immunohistochemistry, we failed to demonstrate any staining of the renal and splenic tissue sections employing an antibody against the decorin core protein.