Sulphated and undersulphated heparan sulphate proteoglycans in a Chinese hamster ovary cell mutant defective in N-sulphotransferase.
The Chinese hamster ovary cell mutant, pgsE-606, synthesizes undersulphated heparan sulphate glycosaminoglycans because of a deficiency in N-sulphotransferase activity [Bame and Esko (1989) J. Biol. Chem. 264, 8059-8065]. We compared the heparan sulphate proteoglycans synthesized by mutant and wild-type cells to determine what effect the undersulphation defect had on proteoglycan structure. The majority of heparan sulphate proteoglycans synthesized by pgsE-606 were undersulphated, but the mutant also synthesized a population of proteoglycans that were sulphated to the same extent as wild-type molecules. Anion-exchange analysis of the glycosaminoglycans in each proteoglycan population showed that they were all modified in the same way. The length of the glycosaminoglycans in each proteoglycan population were similar, suggesting that N-sulphation does not affect chain polymerization. To examine whether the sulphation state of the attached heparan sulphate glycosaminoglycans was dependent on the protein core, we purified syndecan-1 from mutant and wild-type cells using antibodies against the core protein. As with the unfractionated heparan sulphate proteoglycans, pgsE-606 synthesized both undersulphated and sulphated syndecan-1. Each pool contained either undersulphated or sulphated glycosaminoglycan chains respectively. Thus the modification of all heparan sulphate chains on a core protein occurs on a proteoglycan-wide basis (i.e. to the same extent).
Bame
K J
KJ
Department of Biochemistry and Molecular Genetics, School of Medicine, University of Alabama at Birmingham 35294.
Zhang
L
L
David
G
G
Esko
J D
JD
D000818:Animals; D016466:CHO Cells; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D002999:Clone Cells; D006224:Cricetinae; D005944:Glucosamine / Q000378:metabolism; D006025:Glycosaminoglycans / Q000096:biosynthesis / Q000302:isolation & purification; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000096:biosynthesis* / Q000302:isolation & purification; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000302:isolation & purification; D009154:Mutation; D011509:Proteoglycans / Q000096:biosynthesis* / Q000302:isolation & purification; D013431:Sulfates / Q000378:metabolism; D015238:Sulfotransferases / Q000235:genetics*; D013462:Sulfur Radioisotopes; D053667:Syndecans; D014316:Tritium
D005944:Glucosamine; D006025:Glycosaminoglycans; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D008562:Membrane Glycoproteins; D011509:Proteoglycans; D013431:Sulfates; D015238:Sulfotransferases; D013462:Sulfur Radioisotopes; D053667:Syndecans; D014316:Tritium