A novel 4-methylumbelliferyl-beta-D-xyloside derivative, sulfate-O-3-xylosylbeta1-(4-methylumbelliferone), isolated from culture medium of human skin fibroblasts, and its role in methylumbelliferone-initiated glycosaminoglycan biosynthesis.
Human skin fibroblasts were incubated in the presence of 4-methylumbelliferyl-beta-D-xyloside (Xyl-MU). The culture medium was recovered and Xyl-MU derivatives which were initiated by the Xyl-MU acting as a primer were purified. As a result, a novel Xyl-MU derivative was isolated, in addition to previously reported Xyl-MU derivatives such as glycosaminoglycan-MU, Gal-Gal-Xyl-MU, Gal-Xyl-MU, SA-Gal-Xyl-MU, Xyl-Xyl-MU, GlcA-Xyl-MU, and sulfate-GlcA-Xyl-MU. This Xyl-MU derivative was subjected to carbohydrate composition analysis, enzyme digestion, ion-spray mass spectrometric analysis, and Smith degradation. The results indicated that it was sulfate- O -3-Xyl-MU. When Xyl-MU was incubated with [35S]PAPS using a homogenate prepared from the same cultured skin fibroblasts, [35S]sulfate- O -3-Xyl-MU was produced. Moreover, when Xyl-MU was incubated with UDP-[3H]Gal, [3H]galactose was transferred to Xyl-MU, but when sulfate- O -3-Xyl-MU was incubated with UDP-[3H]Gal, [3H]galactose was not transferred. These results indicate that chain elongation from Xyl-MU is inhibited by sulfation of Xyl-MU, and that Xyl-MU sulfation is involved in the control of Xyl-MU-initiated glycosaminoglycan biosynthesis.
Department of Biochemistry and Second Department of Surgery, Hirosaki University School of Medicine, Hirosaki 036-8562, Japan.
T
T
Tazawa
K
K
Takagaki
H
H
Matsuya
T
T
Nakamura
M
M
Sasaki
M
M
Endo
D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006025:Glycosaminoglycans / Q000096:biosynthesis*; D006801:Humans; D006923:Hymecromone / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000378:metabolism*; D013058:Mass Spectrometry; D012867:Skin / Q000166:cytology / Q000378:metabolism*; D014994:Xylose / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000378:metabolism
D006025:Glycosaminoglycans; C114159:sulfate-O-3-xylosyl-(4-methylumbelliferone); D006923:Hymecromone; D014994:Xylose