Effects of detergent on the sulphation of chondroitin by cell-free preparations from chick-embryo epiphyseal cartilage.
The effects of the non-ionic detergent Triton X-100 on 6-sulphation of two species of endogenous nascent proteochondroitin by a chick-embryo cartilage microsomal system was examined. Sulphation of the larger (Type I) species with adenosine 3'-phosphate 5'-phosphosulphate was slightly diminished when Triton X-100 was present, whereas sulphation of the smaller (Type II) species was slightly enhanced. An ordered rather than random pattern of sulphation was obtained for the smaller proteoglycan, but with a considerably lower degree of sulphation than that of the larger proteochondroitin. These differences were consistent with other differences between these two species as described previously. Sulphation of exogenous [14C]chondroitin and exogenous proteo[3H]chondroitin by the microsomal system with Triton X-100 present produced ordered rather than random sulphation patterns. When a 100,000 g supernatant fraction was utilized for sulphation of [14C]chondroitin or proteo[3H]chondroitin, Triton X-100 was not needed, and ordered sulphation was still obtained. When hexasaccharide was used, sulphation of multiple N-acetylgalactosamine residues of the individual hexasaccharides resulted. This was relatively independent of Triton X-100 or the concentration of the hexasaccharide acceptors. With soluble enzyme, sulphation of multiple N-acetylgalactosamine residues on the individual hexasaccharide molecules was even greater, so that tri-sulphated products were found. This suggests that ordered rather than random sulphation of chondroitin with these enzyme preparations is due to enzyme-substrate interaction rather than to membrane organization.
Sugumaran
G
G
Connective Tissue Research Laboratory, Veterans Administration Outpatient Clinic, Bedford, MA 01730.
Silbert
J E
JE
D000818:Animals; D002474:Cell-Free System; D002642:Chick Embryo; D002807:Chondroitin / Q000096:biosynthesis / Q000378:metabolism*; D002809:Chondroitin Sulfates / Q000096:biosynthesis; D002848:Chromatography, DEAE-Cellulose; D003902:Detergents / Q000494:pharmacology*; D006132:Growth Plate / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology*; D013464:Sulfuric Acids / Q000378:metabolism*
D003902:Detergents; D013464:Sulfuric Acids; D011092:Polyethylene Glycols; D017830:Octoxynol; D002807:Chondroitin; D002809:Chondroitin Sulfates