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Title: Differentiation of 3T3-L1 preadipocytes with 3-isobutyl-1-methylxanthine and dexamethasone stimulates cell-associated and soluble chondroitin 4-sulfate proteoglycans

The proteoglycans (cell-associated and culture media) in 3T3-L1 preadipocytes in culture were analyzed before and during differentiation into adipocytes. Cells were metabolically labeled with (35S)sulfate and (3H) glucosamine for 24 h and then extracted and analyzed. There was a 1.68 {plus minus} 0.07-fold increase in the 35S in medium proteoglycan during differentiation, whereas cell-associated proteoglycan radioactivity showed no increase. Analyses of radiolabeled molecules using ion-exchange chromatography, gel filtration, and high performance liquid chromatography after enzymatic or alkaline digestion indicated that all of the 35S label was recovered as two major species of chondroitin 4-sulfate proteoglycans (CSPG-I and CSPG-II) and 7% as heparan sulfate proteoglycan. CSPG-I has a mass of {approximately} 970 kDa with multiple chondroitin sulfate chains (average of 50 kDa each) and a core protein of {approximately} 370 kDa including oligosaccharides. CSPG-II has a mass of 140 kDa with one or two chondroitin sulfate chains (average of 68 kDa each) and a core protein of 41 kDa including oligosaccharides. CSPG-I appears to be similar to versican, whereas CSPG-II is similar to decorin and/or biglycan, found in other fibroblastic cells. Cell differentiation was associated with a specific increase in CSPG-I (4.0 {plus minus} 0.2-fold in media and 3.2 {plus minus}more » 0.5-fold in the cell-associated form). This system should facilitate study of the functional roles of proteoglycans during growth and differentiation.« less
Authors:
; ; ; ;  [1]
  1. (Laboratory of Theoretical and Physical Biology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland (USA))
Publication Date:
OSTI Identifier:
5510089
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Biological Chemistry; (United States); Journal Volume: 266:17
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; FAT CELLS; CELL DIFFERENTIATION; GLYCOPROTEINS; CHEMICAL COMPOSITION; ADIPOSE TISSUE; DEXAMETHASONE; DISACCHARIDES; GLUCOSAMINE; ION EXCHANGE CHROMATOGRAPHY; LIQUID COLUMN CHROMATOGRAPHY; MICE; MOLECULAR WEIGHT; SULFATES; SULFUR 35; TRITIUM; ADRENAL HORMONES; AMINES; ANIMAL CELLS; ANIMAL TISSUES; ANIMALS; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; BODY; CARBOHYDRATES; CHROMATOGRAPHY; CONNECTIVE TISSUE; CONNECTIVE TISSUE CELLS; CORTICOSTEROIDS; DAYS LIVING RADIOISOTOPES; EVEN-ODD NUCLEI; GLUCOCORTICOIDS; HEXOSAMINES; HEXOSES; HORMONES; HYDROGEN ISOTOPES; HYDROXY COMPOUNDS; ISOTOPES; KETONES; LIGHT NUCLEI; MAMMALS; MONOSACCHARIDES; NUCLEI; ODD-EVEN NUCLEI; OLIGOSACCHARIDES; ORGANIC COMPOUNDS; OXYGEN COMPOUNDS; PREGNANES; PROTEINS; RADIOISOTOPES; RODENTS; SACCHARIDES; SEPARATION PROCESSES; SOMATIC CELLS; STEROIDS; SULFUR COMPOUNDS; SULFUR ISOTOPES; TISSUES; VERTEBRATES; YEARS LIVING RADIOISOTOPES 550201* -- Biochemistry-- Tracer Techniques