Both heparin and chondroitin sulfate proteoglycans are intracellular components of dog mastocytoma cell lines
Intracellular proteoglycans (PG), putative markers of mast cell subtype, were isolated and characterized from three dog mastocytome cell lines. Mastocytoma cells were adapted to serial passage as subcutaneous tumors in BALB/c nude mice for 10 generations. Cells were disaggregated and labeled with (number/sup 5/S)SO/sub 4/ in culture. Intracellular /sup 35/S-labeled PG were isolated by sonication in 4M guanidine containing protease and glycosidase inhibitors, then chromatographed on Sepharose CL-4B. All 3 cell lines yielded a major /sup 35/S component at the Vo (Mr greater than or equal to 10/sup 6/). Excluded fractions were treated with alkaline-borohydride and the liberated glycosaminoglycans (GAG) were analyzed on Sephacryl S-200. GAG chains had an apparent Mr of 40-50,000. GAG chains were subjected to chondroitinase ABC and nitrous acid degradation and the products were analyzed on Sepharcyl S-200. In one cell line (BR), 75% of /sup 35/S-labeled GAG was resistant to chondroitinase treatment, but was degraded by nitrous acid, characteristic of heparin. The remaining 25% of /sup 35/S-labeled GAG was degraded to oligosaccharides by chondroitinase indicating chondroitin sulfate (CS). A second cell line (G) also contained both heparin and CS-PG. The remaining cell line (C2) contained 2% heparin-PG and 98% CS-PG. The results demonstrate the simultaneous presence of both heparin and CS-PG in mast cells of clonal origin, and raise important questions on the role of PGs in mast cell granule storage and release mechanisms.
- Research Organization:
- Univ. of California, San Francisco
- OSTI ID:
- 6374458
- Report Number(s):
- CONF-870644-
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 46:6; ISSN FEPRA
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
AMINES
ANIMAL CELLS
ANIMALS
ANTICOAGULANTS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOHYDRATES
CHEMICAL COMPOSITION
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DOGS
DRUGS
EVEN-ODD NUCLEI
GLUCOPROTEINS
HEMATOLOGIC AGENTS
HEPARIN
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MAMMALS
MAST CELLS
METABOLISM
MUCOPOLYSACCHARIDES
NUCLEI
OLIGOSACCHARIDES
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
OXYGEN COMPOUNDS
POLYSACCHARIDES
PROTEINS
RADIOISOTOPES
SACCHARIDES
SOMATIC CELLS
SULFATES
SULFUR 35
SULFUR COMPOUNDS
SULFUR ISOTOPES
TRACER TECHNIQUES
TUMOR CELLS
VERTEBRATES