Comparison of heparan sulfate proteoglycans from equine and human glomerular basement membranes.
1. Proteoglycans extracted from human and equine glomerular basement membranes (GBM) were purified by ion-exchange chromatography and gel filtration. 2. The glycoconjugates had an apparent molecular mass of 200-400 kDa and consisted of 75% protein and 25% glycosaminoglycan. Glycosidase and HNO2 treatment and the amino sugar and sulfate composition of both proteoglycan preparations identified heparan sulfate (HS) as the predominant saccharide chain. 3. Hydrolysis with trifluoromethanesulfonic acid yielded comparable core proteins with molecular masses of ca 160 and 120 kDa. 4. The HS chains had an apparent molecular mass of 18 kDa. Results of heparitinase digestion and HNO2-treatment indicated a clustering of sulfate groups in the distal part of the HS side chains. 5. Peptide mapping after trypsin, clostripain or V8 protease digestion of radiolabeled human and equine heparan sulfate proteoglycans (HSPG) preparations with three different separation techniques showed large differences. 6. Polyclonal antisera raised against the HSPGs reacted against the core proteins. Both HSPG preparations and their antisera showed ca 40% cross-reactivity. About 50% of monoclonal antisera elicited against one HSPG preparation showed reaction with both HSPG preparations. 7. Polyclonal antisera stained all basement membranes in an intense linear fashion in indirect immunofluorescence studies of kidney sections from horse, man and various mammalian species. 8. Biochemical and immunological data indicate that HSPGs from equine and human GBM have a comparable structure, but the core proteins differ considerably.
van den Heuvel
L P
LP
Department of Biochemistry, University of Nijmegen, The Netherlands.
van den Born
J
J
Veerkamp
J H
JH
Janssen
G H
GH
van de Velden
T J
TJ
Monnens
L A
LA
Schröder
C H
CH
Berden
J H
JH
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001485:Basement Membrane / Q000737:chemistry*; D002241:Carbohydrates / Q000032:analysis; D011508:Chondroitin Sulfate Proteoglycans / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D000939:Epitopes / Q000276:immunology; D005455:Fluorescent Antibody Technique; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D006736:Horses / Q000378:metabolism*; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D015151:Immunoblotting; D007678:Kidney Glomerulus / Q000737:chemistry*; D008698:Mesylates; D008970:Molecular Weight; D010449:Peptide Mapping; D011133:Polysaccharide-Lyases / Q000378:metabolism
D000596:Amino Acids; D002241:Carbohydrates; D011508:Chondroitin Sulfate Proteoglycans; D000939:Epitopes; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D007106:Immune Sera; D008698:Mesylates; D011133:Polysaccharide-Lyases; C021349:heparitinsulfate lyase; C012077:trifluoromethanesulfonic acid