title: Distribution in cesium chloride gradients of proteoglycans of chick embryo brain and characterization of a large aggregating proteoglycan.
abstract: Proteoglycans were extracted from 14-day chick embryo brains, which had been labelled in vitro with [35S]sulfate or 3H-labelled amino acids. 4.0 M guanidinium chloride (containing proteinase inhibitors) extracted 94% of the 35S-labelled glycoconjugates. Following cesium chloride equilibrium centrifugation, the proteoglycans in each fraction were characterized by chromatography on Sepharose CL-2B. The most dense fraction (D1), which contained no detectable non-proteoglycan proteins, contained a large, aggregating chondroitin sulfate proteoglycan in addition to small chondroitin sulfate and heparan sulfate proteoglycans. The less dense fractions (D2-D6) contained both small chondroitin sulfate and heparan sulfate proteoglycans. Removal of hyaluronate from the D1 sample by digestion with Streptomyces hyaluronidase in the presence of proteinase inhibitors showed that aggregation of the large chondroitin sulfate proteoglycan is hyaluronate-dependent. Aggregation was restored by re-addition of hyaluronate. Reduction and alkylation, which blocked aggregation of a cartilage A1 proteoglycan, did not interfere with aggregation of the large brain proteoglycan.
authors.affiliation: Department of Zoology, University of British Columbia, Vancouver, Canada.
authors.forename: T
authors.initials: T
authors.lastname: Crawford
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D002499:Centrifugation, Density Gradient; D002586:Cesium; D002642:Chick Embryo; D002712:Chlorides; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006025:Glycosaminoglycans / Q000302:isolation & purification; D046911:Macromolecular Substances; D011509:Proteoglycans / Q000302:isolation & purification* / Q000378:metabolism; D013462:Sulfur Radioisotopes; D014316:Tritium
chemical_list: D002712:Chlorides; D006025:Glycosaminoglycans; D046911:Macromolecular Substances; D011509:Proteoglycans; D013462:Sulfur Radioisotopes; D014316:Tritium; D002586:Cesium; C028019:cesium chloride