Proteoglycan biosynthesis in murine monocytic leukemic (M1) cells before and after differentiation
- National Institute of Dental Research, Bethesda, MD (USA)
Murine monocytic leukemic (M1) cells were cultured in the presence of ({sup 3}H)glucosamine and ({sup 35}S)sulfate. Labeled proteoglycans were purified by anion exchange chromatography and characterized by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with chemical and enzymatic degradation. M1 cells synthesize a single predominant species of proteoglycan which distributes almost equally between the cell and medium after 17 h labeling. The cell-associated proteoglycan has an overall size of about 135 kDa and contains three to five chondroitin sulfate chains (28-31 kDa each) attached to a chondroitinase-generated core protein of 28 kDa. The synthesis and subsequent secretion of this proteoglycan was enhanced 4-5-fold in cells induced to differentiate into macrophages. This was not a phenomenon of arrest in the G0/G1 stage of the cell cycle, since density inhibited undifferentiated cells arrested at this stage did not increase proteoglycan synthesis. The chondroitin sulfate chains contained exclusively chondroitin 4- and 6-sulfate; however, the ratio of these two disaccharides differed between the medium- and cell-associated proteoglycans, and changed during progression of the cells into a fully differentiated phenotype. Pulse-chase kinetics indicate the presence of two distinct pools of proteoglycan; one that is secreted very rapidly from the cell after a approximately 1-h lag, and a second pool that is turned over in the cell with a half-time of approximately 3.5 h. Subtle differences in the glycosylation patterns of the medium- and cell-associated species are consistent with synthesis of two pools. Papain digestion suggests that the chondroitin sulfate chains are clustered on a small protease resistant peptide. The data suggest that this proteoglycan is similar to the serglycin proteoglycan family.
- OSTI ID:
- 5607776
- Journal Information:
- Journal of Biological Chemistry; (USA), Vol. 264:22; ISSN 0021-9258
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
GLYCOPROTEINS
BIOSYNTHESIS
TUMOR CELLS
CELL DIFFERENTIATION
CELL CYCLE
ELECTROPHORESIS
FRACTIONATION
GLUCOSAMINE
HYDROLYSIS
ION EXCHANGE CHROMATOGRAPHY
LEUKEMIA
MACROPHAGES
MICE
MOLECULAR WEIGHT
PAPAIN
PEPTIDE HYDROLASES
PHENOTYPE
PROTEIN STRUCTURE
SULFATES
SULFUR 35
TRACER TECHNIQUES
TRITIUM COMPOUNDS
AMINES
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOHYDRATES
CHEMICAL REACTIONS
CHROMATOGRAPHY
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DECOMPOSITION
DISEASES
ENZYMES
EVEN-ODD NUCLEI
HEMIC DISEASES
HEXOSAMINES
HEXOSES
HYDROGEN COMPOUNDS
HYDROLASES
IMMUNE SYSTEM DISEASES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LYSIS
MAMMALS
MONOSACCHARIDES
NEOPLASMS
NUCLEI
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PHAGOCYTES
PROTEINS
RADIOISOTOPES
RODENTS
SACCHARIDES
SEPARATION PROCESSES
SH-PROTEINASES
SOLVOLYSIS
SOMATIC CELLS
SULFUR COMPOUNDS
SULFUR ISOTOPES
SYNTHESIS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques