Isolation and partial characterization of heparan sulphate proteoglycan from the human glomerular basement membrane
| Autor: | J. van den Born, T.J.A.M. van de Velden, C. H. Schröder, L.P.W.J. van den Heuvel, Leo A. H. Monnens, J.H. Veerkamp, Jhm Berden |
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| Přispěvatelé: | Groningen Kidney Center (GKC), Groningen Institute for Organ Transplantation (GIOT), Guided Treatment in Optimal Selected Cancer Patients (GUTS) |
| Rok vydání: | 1989 |
| Předmět: |
Kidney Cortex
Renal glomerulus Glycoconjugate Kidney Glomerulus Blotting Western Fluorescent Antibody Technique Enzyme-Linked Immunosorbent Assay Kidney Cortex/cytology Amino Acids/analysis Biochemistry Basement Membrane Proteoglycans/isolation & purification Basement Membrane/analysis Heparitin Sulfate/isolation & purification medicine Humans Amino Acids Molecular Biology Glycosaminoglycans Alanine Antiserum Basement membrane chemistry.chemical_classification Molecular mass biology Chemistry Chondroitin Sulfate Proteoglycans/isolation & purification Kidney Glomerulus/analysis Glomerular basement membrane Glycosaminoglycans/isolation & purification Hexosamines Cell Biology Chromatography Ion Exchange Hexosamines/analysis Molecular Weight medicine.anatomical_structure Chondroitin Sulfate Proteoglycans Proteoglycan Chromatography Gel biology.protein Proteoglycans Electrophoresis Polyacrylamide Gel Heparitin Sulfate Heparan Sulfate Proteoglycans Research Article |
| Zdroj: | Biochemical Journal, 264(2), 457-465. PORTLAND PRESS LTD |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bj2640457 |
| Popis: | Heparan sulphate proteoglycan was solubilized from human glomerular basement membranes by guanidine extraction and purified by ion-exchange chromatography and gel filtration. The yield of proteoglycan was approx. 2 mg/g of basement membrane. The glycoconjugate had an apparent molecular mass of 200-400 kDa and consisted of about 75% protein and 25% heparan sulphate. The amino acid composition was characterized by a high content of glycine, proline, alanine and glutamic acid. Hydrolysis with trifluoromethanesulphonic acid yielded core proteins of 160 and 110 kDa (and minor bands of 90 and 60 kDa). Alkaline NaBH4 treatment of the proteoglycan released heparan sulphate chains with an average molecular mass of 18 kDa. HNO2 oxidation of these chains yielded oligosaccharides of about 5 kDa, whereas heparitinase digestion resulted in a more complete degradation. The data suggest a clustering of N-sulphate groups in the peripheral regions of the glycosaminoglycan chains. A polyclonal antiserum raised against the intact proteoglycan showed reactivity against the core protein. It stained all basement membranes in an intense linear fashion in immunohistochemical studies on frozen kidney sections from man and various mammalian species. |
| Databáze: | OpenAIRE |
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