Secretion of chondroitin 6-sulfotransferase and chondroitin 4-sulfotransferase from cultured chick embryo chondrocytes.
We found that chondroitin 6-sulfotransferase and chondroitin 4-sulfotransferase were released into the culture medium from the cultured chick embryo chondrocytes. Since the release of the sulfotransferases was observed not only in serum-supplemented medium but also in serum-free medium, the released sulfotransferases were unlikely to be derived from serum. Addition of ascorbate to the serum-free medium supported the continuous release of the sulfotransferases. Monensin, which is known to cause dilatation of the Golgi apparatus and to inhibit sulfation of proteoglycan, was found to affect the release of the sulfotransferases. In the presence of 10(-6) M monensin, chondroitin 6-sulfotransferase activity in the cell layer was decreased to less than one tenth of the control, and the rate of the release of the activity became much smaller than the control after the initial rapid release. The activity of chondroitin 4-sulfotransferase was also affected by monensin, but the reduction of the chondroitin 4-sulfotransferase activity in the cell layer was not so great as the reduction of chondroitin 6-sulfotransferase activity. Unlike to the microsomal sulfotransferases, both chondroitin 6-sulfotransferase and chondroitin 4-sulfotransferase released into the culture medium were retained in the soluble fraction after centrifugation at 100,000 x g for 60 min, and were not activated by detergent. pH optimum and requirements for sulfhydryl compounds of the released sulfotransferases were similar to those observed previously in the chondroitin sulfotransferases from chick embryo cartilage and from cultured chick embryo chondrocytes. These results suggest that chondroitin sulfotransferases, which are localized in the Golgi apparatus, may be secreted to the extracellular space in a soluble form under the culture conditions.
Habuchi
O
O
Department of Chemistry, Aichi Kyoiku University, Kariya, Japan.
Tsuzuki
M
M
Takeuchi
I
I
Hara
M
M
Matsui
Y
Y
Ashikari
S
S
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D002478:Cells, Cultured; D002642:Chick Embryo; D016895:Culture Media, Serum-Free; D006614:Hindlimb / Q000033:anatomy & histology / Q000166:cytology / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D008985:Monensin / Q000494:pharmacology; D012995:Solubility; D013438:Sulfhydryl Compounds; D015238:Sulfotransferases / Q000378:metabolism*; D013643:Tarsus, Animal / Q000166:cytology / Q000201:enzymology; D013977:Tibia / Q000166:cytology / Q000201:enzymology; D000097811:Carbohydrate Sulfotransferases
D016895:Culture Media, Serum-Free; D008985:Monensin; D013438:Sulfhydryl Compounds; D015238:Sulfotransferases; D000097811:Carbohydrate Sulfotransferases; C019893:chondroitin 4-sulfotransferase