A rat osteogenic cell line (UMR 106-01) synthesizes a highly sulfated form of bone sialoprotein
- National Institutes of Health, Bethesda, MD (USA)
The rat osteosarcoma cell line (UMR 106-01) synthesizes and secretes relatively large amounts of a sulfated glycoprotein into its culture medium (approximately 240 ng/10(6) cells/day). This glycoprotein was purified, and amino-terminal sequence analysis identified it as bone sialoprotein (BSP). (35S)Sulfate, (3H)glucosamine, and (3H)tyrosine were used as metabolic precursors to label the BSP. Sulfate esters were found on N- and O-linked oligosaccharides and on tyrosine residues, with about half of the total tyrosines in the BSP being sulfated. The proportion of 35S activity in tyrosine-O-sulfate (approximately 70%) was greater than that in N-linked (approximately 20%) and O-linked (approximately 10%) oligosaccharides. From the deduced amino acid sequence for rat BSP, the results indicate that on average approximately 12 tyrosine residues, approximately 3 N-linked, and approximately 2 O-linked oligosaccharides are sulfated/molecule. The carboxyl-terminal quarter of the BSP probably contains most, if not all, of the sulfated tyrosine residues because this region of the polypeptide contains the necessary requirements for tyrosine sulfation. Oligosaccharide analyses indicated that for every N-linked oligosaccharide on the BSP, there are also approximately 2 hexa-, approximately 5 tetra-, and approximately 2 trisaccharides O-linked to serine and threonine residues. On average, the BSP synthesized by UMR 106-01 cells would contain a total of approximately 3 N-linked and approximately 25 of the above O-linked oligosaccharides. This large number of oligosaccharides is in agreement with the known carbohydrate content (approximately 50%) of the BSP.A
- OSTI ID:
- 7009598
- Journal Information:
- Journal of Biological Chemistry; (USA), Vol. 265:9; ISSN 0021-9258
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
GLYCOPROTEINS
BIOSYNTHESIS
AMINO ACID SEQUENCE
CHROMATOGRAPHY
GLUCOSAMINE
OLIGOSACCHARIDES
OSTEOSARCOMAS
PRECURSOR
RATS
SECRETION
SULFATES
SULFUR 35
TRACER TECHNIQUES
TRITIUM COMPOUNDS
TUMOR CELLS
TYROSINE
AMINES
AMINO ACIDS
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOHYDRATES
CARBOXYLIC ACIDS
DAYS LIVING RADIOISOTOPES
DISEASES
EVEN-ODD NUCLEI
HEXOSAMINES
HEXOSES
HYDROGEN COMPOUNDS
HYDROXY ACIDS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MAMMALS
MOLECULAR STRUCTURE
MONOSACCHARIDES
NEOPLASMS
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PROTEINS
RADIOISOTOPES
RODENTS
SACCHARIDES
SARCOMAS
SEPARATION PROCESSES
SKELETAL DISEASES
SULFUR COMPOUNDS
SULFUR ISOTOPES
SYNTHESIS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques
550901 - Pathology- Tracer Techniques