Purification form human hepatoma cells of a 130-kDa membrane glycoprotein with properties of the heparin-binding growth factor receptor.
The detergent-soluble 125I-labeled receptor complex resulting after affinity cross-linking of 125I-heparin-binding growth factor type one (HBGF-1, m = 15.2-kDa) to HepG2 cells had an apparent molecular mass of 145-kDa, eluted from immobilized wheat germ lectin in the presence of N-acetylglucosamine, shifted to apparent mass of 128-kDa when treated with N-glycanase and shifted to apparent mass of 205-kDa after reduction, carboxymethylation and succinylation. Electrophoretic analysis of HepG2 cell membrane proteins revealed a major silver-stained protein of apparent molecular mass of 130-kDa that has correlative properties. These properties were used to purify the 130-kDa HepG2 glycoprotein to apparent homogeneity and suggest the glycoprotein as a candidate for the human HBGF receptor.
DiSorbo
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W. Alton Jones Cell Science Center, Inc., Lake Placid, New York 12946.
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McKeehan
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D000117:Acetylglucosamine; D006528:Carcinoma, Hepatocellular / Q000032:analysis*; D055598:Chemical Phenomena; D002621:Chemistry; D002846:Chromatography, Affinity; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D016220:Fibroblast Growth Factor 1; D006026:Glycoside Hydrolases; D006133:Growth Substances / Q000378:metabolism; D006493:Heparin / Q000378:metabolism; D006801:Humans; D007457:Iodine Radioisotopes; D008113:Liver Neoplasms / Q000032:analysis*; D008562:Membrane Glycoproteins / Q000302:isolation & purification*; D008934:Mitogens; D008970:Molecular Weight; D011975:Receptors, Mitogen / Q000302:isolation & purification*; D040262:Receptors, Vascular Endothelial Growth Factor; D014407:Tumor Cells, Cultured
D003432:Cross-Linking Reagents; D006133:Growth Substances; D007457:Iodine Radioisotopes; D008562:Membrane Glycoproteins; D008934:Mitogens; D011975:Receptors, Mitogen; D016220:Fibroblast Growth Factor 1; D006493:Heparin; D040262:Receptors, Vascular Endothelial Growth Factor; D006026:Glycoside Hydrolases; C020668:glycanase; D000117:Acetylglucosamine