Biomedical Chromatography · 2001 · 20 citations · 31 references
Life SciencesBiochemistryImmunologyCellular BiochemistryGlycobiologyImmunopathologyChromatographyExtracellular Matrix GlycansGlycan AnalysisGlycan BiosynthesisGlycan StructureLiquid ChromatographyChondroitin Sulfate BiosynthesisGlycosaminoglycan BiosynthesisGlycosaminoglycan StructureGlycosaminoglycansHeparan Sulfate BiosynthesisHeparan Sulfate SynthesisHigh-performance Liquid ChromatographyIon-exchange Liquid Chromatography
Abstract Identification of glycosaminoglycans (GAGs) synthesized by three human leukaemic cell lines—Jurkat (T‐cell leukaemia), Daudi (Burkitt's lymphoma, B‐cell leukaemia) and THP‐1 (acute monocytic leukemia)—and normal peripheral blood mononuclear cells (PBMC) and their distribution among cell membrane and culture medium were studied. GAGs were isolated using ion‐exchange chromatography on DEAE‐Sephacel and their composition and fine chemical structure were studied using high‐performance liquid chromatography with radiochemical detection. All cell lines synthesize chondroitin sulphate (CS) and heparan sulphate (HS) in both cell membrane and culture medium. No hyaluronan was detected using treatment with specific lyases and highly sensitive HPLC methodology. CS is the major secreted GAG in all cell lines tested and the major cell retained GAG in Jurkat and Daudi. HS is the major GAG in the cell membrane of THP‐1. The amounts of distinct GAGs synthesized by all cancer cell lines differ from those produced by normal PBML indicating a major role of GAGs in malignant transformation of human lymphocytes and monocytes. Copyright © 2001 John Wiley & Sons, Ltd. Abbreviations used: CS chondroitin sulphate GAGs glycosaminoglycans HS heparan sulphate PBMC peripheral blood mononuclear cells PG proteoglycans PMSF, phenylmethylsulphonyl fluoride
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