Differential and cell-type specific microheterogeneity of high mannose-type Asn-linked oligosaccharides of human transferrin receptor.
The structural analysis of high mannose-type Asn-linked (N-linked) oligosaccharides of the human transferrin receptor (hTR) from D-[2-3H]mannose metabolic-radiolabeled human cells--A431, K562, BeWo, and HL60--was investigated. The radiolabeled hTR glycopeptides were prepared and fractionated by a lectin chromatography of Concanavalin A-Sepharose. The composition analysis of hTR glycopeptides revealed that Con A-I contains both mannose and fucose, whereas Con A-III has mannose exclusively. The Con A-III glycopeptides were treated with Endo H. The released oligosaccharides were charge-fractionated by QAE-Sephadex. The neutral oligosaccharides were further size-fractionated by an amine absorption high performance liquid chromatography (HPLC). Our results demonstrate that the high mannose-type oligosaccharides of hTR ranged in size from Man5-R to Man9-R with cell-type specific patterns. A relative amount of each component was found to be differentially heterogeneous among the four different human cell lines.
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Korea Research Institute of Bioscience and Biotechnology (KRIBB), Cell Biology Division, Animal Cell and Medical Glycobiology Laboratory, Taejon.
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D001216:Asparagine / Q000737:chemistry; D002460:Cell Line; D002847:Chromatography, Agarose; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D018922:HL-60 Cells; D006801:Humans; D020014:K562 Cells; D008358:Mannose / Q000737:chemistry; D015394:Molecular Structure; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification; D011990:Receptors, Transferrin / Q000737:chemistry*
D009844:Oligosaccharides; D011990:Receptors, Transferrin; D001216:Asparagine; D008358:Mannose