PMID: 8477709

 

    Legend: Gene, Sites

Title : Structures of sialylated oligosaccharides of human erythropoietin expressed in recombinant BHK-21 cells

Abstract :
  1. The native structures of the Asn-linked oligosaccharides and the O-glycans at Ser126 of human erythropoietin expressed from recombinant BHK cells have been elucidated
  2. Enzymatically released N-glycans were studied by methylation analyses, fast-atom-bombardment mass spectrometry as well as one- and two-dimensional 1H-NMR spectrometry at 600 MHz
  3. Many (82.7%) were found to be tetraantennary N-acetyllactosamine-type (22.8% with one, 3.6% with two and 0.4% with three N-acetyllactosamine repeats) being tetrasialylated (41%), trisialylated (29.6%) and disialylated (12.2%)
  4. A few (9.7%; 4.1% 2,4-branched, 5.6%, 2,6-branched) of the chains were triantennary (5.4% trisialyl, 4.3% disialyl) and 4.6% were of the disialyl diantennary type
  5. Almost all of the innermost GlcNAc residues were alpha 1-6 fucosylated and NeuAc was exclusively alpha 2-3 linked to Gal beta 1-4GlcNAc-R; 60% of the protein was found to be O-glycosylated at Ser126 ; structures were monosialylated (70%) or disialylated (30%) forms of the Gal beta 1-3GalNAc core type
  6. Glycosylation patterns at individual Asn-Xaa-Thr/Ser sites were determined by analytical high-pH anion-exchange chromatography with pulsed amperometric detection
  7. Only tetraantennary chains with 0-3 N-acetyllactosamine repeats were detected at Asn38 and Asn83 , while almost all of the di- and triantennary oligosaccharides were attached to Asn24
  8. Batch analysis of different preparations of recombinant erythropoietin revealed the high reproducibility of the production procedure
  9. Structures containing terminal GalNAc-GlcNAc were detected in small amounts in a few batches
Output (sent_index, trigger, protein, sugar, site):
  • 0. erythropoietin, , erythropoietin, sialylated oligosaccharides, -
  • 0. sialylated, , -, sialylated oligosaccharides, -
  • 1. Asn-linked, , -, the Asn-linked oligosaccharides, Asn
  • 1. Ser126, , -, the O-glycans, Ser126
  • 5. O-glycosylated, , protein, -, Ser126
  • 5. disialylated, , -, monosialylated (70%) or disialylated (30%) forms, -
  • 5. disialylated, , -, the Gal beta 1-3GalNAc core type, -
  • 5. fucosylated, , -, all, -
  • 5. fucosylated, , -, the innermost GlcNAc residues, -
  • 5. monosialylated, , -, monosialylated (70%) or disialylated (30%) forms, -
  • 5. monosialylated, , -, the Gal beta 1-3GalNAc core type, -
  • 7. attached, , -, all, Asn24
  • 7. attached, , -, di-, Asn24
  • 7. attached, , -, triantennary oligosaccharides, Asn24
  • 7. detected, , -, Only tetraantennary chains, Asn38 and Asn83
Output(Part-Of) (sent_index, protein, site):
  • 1. erythropoietin, Ser126
*Output_Site_Fusion* (sent_index, protein, sugar, site):
  • 1. erythropoietin, the O-glycans, Ser126
  • 5. protein, -, Ser126
  • 7. erythropoietin, Only tetraantennary chains, Asn38 and Asn83
  • 7. erythropoietin, all, Asn24
  • 7. erythropoietin, di-, Asn24
  • 7. erythropoietin, triantennary oligosaccharides, Asn24

 

 

Protein NCBI ID SENTENCE INDEX
erythropoietin 2056 0,1,8