Purification and partial characterization of glycosaminoglycans and proteoglycans from cultured rabbit smooth muscle cells
Glycosaminoglycans synthesized by cultured rabbit smooth muscle cells were isolated after incorporation of (/sup 3/H)-glucosamine into glycosaminoglycans in the presence or absence of 10% fetal bovine serum. Glycosaminoglycans were quantitated by two-dimensional electrophoresis after proteolytic digestion of the cell layers and media. The results show that the presence of serum has no effect on the chondroitin sulfate, heparan sulfate and dermatan sulfate content of the cell layers. The incorporation of (/sup 3/H)-glucosamine into hyaluronic acid of the cell layers was three times higher in the presence of serum. In the medium , the quantity of hyaluronic was two times higher in the presence of serum while the other glycosaminoglycans remained unchanged. The incorporation of (/sup 3/H)-glucosamine into hyaluronic acid was unaffected by the presence of serum. Specific proteoglycans were isolated from medium after with (/sup 35/S)-sulfate and (/sup 3/H)-serine by isopycnic ultracentrifugation and chromatography on Sepharose CL-4B and DEAE-cellulose. Preparations contained a chondroitin sulfate proteoglycan, a condroitin sulfate-dermatan sulfate proteoglycan and a heparan sulfate proteoglycan. Glycosaminoglycans and proteoglycans synthesized by rabbit aorta smooth muscle cells are similar to those from human aorta.
- Research Organization:
- Boston Univ., MA (USA)
- OSTI ID:
- 5073297
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
GLUCOPROTEINS
BIOSYNTHESIS
HEXOSAMINES
MUSCLES
BIOCHEMICAL REACTION KINETICS
ANIMAL CELLS
BLOOD SERUM
CELL CULTURES
ELECTROPHORESIS
GLUCOSAMINE
ION EXCHANGE CHROMATOGRAPHY
LABELLED COMPOUNDS
RABBITS
SULFUR 35
TRITIUM COMPOUNDS
AMINES
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOHYDRATES
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
EVEN-ODD NUCLEI
HEXOSES
ISOTOPES
KINETICS
LIGHT NUCLEI
MAMMALS
MONOSACCHARIDES
NUCLEI
ORGANIC COMPOUNDS
PROTEINS
RADIOISOTOPES
REACTION KINETICS
SACCHARIDES
SEPARATION PROCESSES
SULFUR ISOTOPES
SYNTHESIS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques
550301 - Cytology- Tracer Techniques