Heparan sulfate proteoglycan from human tubular basement membrane. Comparison with this component from the glomerular basement membrane.
Heparan sulfate proteoglycan (HSPG) was extracted from human tubular basement membrane (TBM) with guanidine and purified by ion-exchange chromatography and gel filtration. The glycoconjugate was sensitive to heparitinase and resistant to chondroitinase ABC, had an apparent molecular mass of 200-400 kDa and consisted of 70% protein and 30% glycosaminoglycan. The amino acid composition was characterized by its high content of glycine, proline, alanine and glutamic acid. Hydrolysis with trifluoromethanesulfonic acid yielded core proteins of 160 and 110 kDa. The heparan sulfate (HS) chains obtained after alkaline NaBH4 treatment had a molecular mass of about 18 kDa. Results of heparitinase digestion and HNO2 treatment suggest a clustering of sulfate groups in the distal portion of the HS side chains. These chemical data are comparable to those obtained previously on glomerular basement membrane (GBM) HSPG (Van den Heuvel et al. (1989) Biochem. J. 264, 457-465). Peptide patterns obtained after trypsin, clostripain or V8 protease digestion of TBM and GBM HSPG preparations showed a large similarity. Polyclonal antisera and a panel of monoclonal antibodies raised against both HSPG preparations and directed against the core protein showed complete cross-reactivity in ELISA and on Western blots. They stained all basement membranes in an intense linear fashion in indirect immunofluorescence studies on human kidneys. Based on these biochemical and immunological data we conclude that HSPGs from human GBM and TBM are identical, or at least very closely related, proteins.
van den Heuvel
L P
LP
Department of Biochemistry, University of Nijmegen, The Netherlands.
van den Born
J
J
Veerkamp
J H
JH
van de Velden
T J
TJ
Schenkels
L
L
Monnens
L A
LA
Schröder
C H
CH
Berden
J H
JH
D000596:Amino Acids / Q000032:analysis; D000911:Antibodies, Monoclonal / Q000276:immunology; D001485:Basement Membrane / Q000032:analysis / Q000276:immunology; D011508:Chondroitin Sulfate Proteoglycans / Q000032:analysis*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005455:Fluorescent Antibody Technique; D006025:Glycosaminoglycans / Q000032:analysis*; D019791:Guanidine; D006146:Guanidines / Q000494:pharmacology; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000032:analysis*; D006801:Humans; D006868:Hydrolysis; D007678:Kidney Glomerulus / Q000032:analysis* / Q000187:drug effects / Q000276:immunology; D007684:Kidney Tubules / Q000032:analysis*; D008698:Mesylates / Q000494:pharmacology; D011133:Polysaccharide-Lyases / Q000494:pharmacology; D011509:Proteoglycans / Q000032:analysis*
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D011508:Chondroitin Sulfate Proteoglycans; D006025:Glycosaminoglycans; D019791:Guanidine; D006146:Guanidines; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D008698:Mesylates; D011133:Polysaccharide-Lyases; D011509:Proteoglycans; C021349:heparitinsulfate lyase; C012077:trifluoromethanesulfonic acid