Title : Comparative structural study of N-linked oligosaccharides of urinary and recombinant erythropoietins
Abstract :
- The structures of the N-linked oligosaccharides of the urinary erythropoietin (u- EPO ) purified from urine of aplastic anemic patients were analyzed and compared with those for recombinant erythropoietin (r- EPO ) prepared with baby hamster kidney ( BHK ) cells
- Asparagine-linked neutral oligosaccharides were released from each EPO protein by N-oligosaccharide glycopeptidase (almond) digestion
- The reducing ends of the oligosaccharide chains thus obtained were aminated with a fluorescent reagent, 2-aminopyridine, and the mixture of pyridylamino derivatives of the oligosaccharides was separated by high-performance liquid chromatography (HPLC) on an ODS silica column
- More than 8 and 13 kinds of oligosaccharide fractions for u- EPO and r- EPO ( BHK ), respectively, were completely separated by the one-step HPLC procedure
- The structure of each oligosaccharide thus isolated was analyzed by a combination of sequential exoglycosidase digestion and another kind of HPLC with an amide-silica column
- Furthermore, high-resolution proton nuclear magnetic resonance (1H NMR) spectroscopy and methylation analyses were carried out in the case of r- EPO ( BHK )
- (ABSTRACT TRUNCATED AT 250 WORDS
Output (sent_index, trigger,
protein,
sugar,
site):
- 1. erythropoietin, , erythropoietin, the N-linked oligosaccharides, -
- 2. Asparagine-linked, , -, Asparagine-linked neutral oligosaccharides, Asparagine
- 2. released, , EPO protein, Asparagine-linked neutral oligosaccharides, -
Output(Part-Of) (sent_index,
protein,
site):
*Output_Site_Fusion* (sent_index,
protein,
sugar,
site):