Comparative structural study of N-linked oligosaccharides of urinary and recombinant erythropoietins
The structures of the N-linked oligosaccharides of the urinary erythropoietin (u-EPOI) 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 (/sup 1/H NMR) spectroscopy and methylation analyses were carried out in the case of r-EPO (BHK).
- Research Organization:
- Snow Brand Milk Products Co. Ltd., Tochigi (Japan)
- OSTI ID:
- 7001556
- Journal Information:
- Biochemistry; (United States), Vol. 27:15
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
OLIGOSACCHARIDES
MOLECULAR STRUCTURE
NMR SPECTRA
ANEMIAS
ANIMAL CELLS
ASPARAGINE
ERYTHROPOIETIN
KIDNEYS
LIQUID COLUMN CHROMATOGRAPHY
METHYLATION
NUCLEAR MAGNETIC RESONANCE
PATIENTS
PROTONS
URINE
AMIDES
AMINO ACIDS
BARYONS
BIOLOGICAL MATERIALS
BIOLOGICAL WASTES
BODY
BODY FLUIDS
CARBOHYDRATES
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
CHROMATOGRAPHY
DISEASES
ELEMENTARY PARTICLES
FERMIONS
HADRONS
HEMIC DISEASES
MAGNETIC RESONANCE
MATERIALS
MITOGENS
NUCLEONS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PROTEINS
RESONANCE
SACCHARIDES
SEPARATION PROCESSES
SPECTRA
SYMPTOMS
WASTES
550601* - Medicine- Unsealed Radionuclides in Diagnostics