[Effect of p-nitrophenyl-xyloside on the biosynthesis of proteoglycan in rat ovarian granulosa cells--analyses of glycosaminoglycan synthesis in the Golgi apparatus].
Biosynthesis of proteoglycans and glycosaminoglycans in the presence of p-nitrophenyl-xyloside was studied using a primary rat ovarian granulosa cell culture system. Addition of p-nitrophenyl-xyloside into cell culture medium caused about a 700% increase of [³⁵S]sulfate incorporation (ED50 at 0.03 mM) into macromolecules, which included free chondroitin sulfate chains initiated on xyloside and native proteoglycans. Free chondroitin sulfate chains initiated on xyloside were almost exclusively secreted into the medium. The molecular size of chondroitin sulfate chains decreased from 40,000 to 21,000 as the total [³⁵S]sulfate incorporation was enhanced, suggesting that enhanced synthesis of chondroitin sulfate perturbed the normal mechanism of glycosaminoglycan chain termination. Biosynthesis of heparan sulfate proteoglycans was reduced by approximately 50%, likely due to competition at the level of UDP-sugar precursors. [³⁵S]Sulfate incorporation was shut down by the addition of cycloheximide with an initial half time of approximately 2 hr in the presence of xyloside, while that in the absence of xyloside was about 20 min. The difference likely reflects the turnover rate of glycosaminoglycan synthesizing capacity as a whole. The turnover rate of glycosaminoglycan synthesizing capacity observed in ovarian granulosa cells was much shorter than that observed in chondrocytes, reflecting the relative dominance of proteoglycan biosynthetic activity in the total metabolic activity of the cells.
Yanagishita
Masaki
M
Biochemistry, Department of Hard Tissue Engineering, Division of Bio-Matrix, Graduate School, Tokyo Medical and Dental University.
Tsuchida
Sachio
S
Yokoyama
Miki
M
Inoue
Katarzyna Anna
KA
D000818:Animals; D005260:Female; D006025:Glycosaminoglycans / Q000096:biosynthesis*; D006027:Glycosides / Q000494:pharmacology*; D006056:Golgi Apparatus / Q000378:metabolism*; D006107:Granulosa Cells / Q000378:metabolism*; D011509:Proteoglycans / Q000096:biosynthesis*; D051381:Rats; D017207:Rats, Sprague-Dawley
D006025:Glycosaminoglycans; D006027:Glycosides; D011509:Proteoglycans; C008646:4-nitrophenyl beta-D-xyloside