1. Fig. 1. Gel-filtration chromatography on Sephadex G-15
Fig. 1. Gel-filtration chromatography on Sephadex G-15. The dialyzable fraction of culture medium was concentrated and applied to a Sephadex G-15 column (4.1 × 150 cm). Elution was performed with distilled water at flow rate of 50 ml/h, and 25 ml fractions were collected. The eluate was monitored with a fluorescence detector at excitation and emission wavelengths of 325 and 380 nm, respectively. The fractions indicated by the horizontal bar were collected and used for further purification.
From: A novel 4-methylumbelliferyl-β-D-xyloside derivative, sulfate-O-3-xylosylβ1-(4-methylumbelliferone), isolated from culture medium of human skin fibroblasts, and its role in methylumbelliferone-initiated glycosaminoglycan biosynthesis
Glycobiology. 1998;8(9): doi: /glycob/
Glycobiology | © 1998 Oxford University Press

2. Fig. 2. Ion-exchange chromatography on DEAE-Sephacel
Fig. 2. Ion-exchange chromatography on DEAE-Sephacel. The fractions recovered from Sephadex G-15 chromatography were applied to a DEAE-Sephacel column (2.3 × 40 cm). Elution was performed with a linear gradient of 0–1.0 M NaCl at a flow rate of 30 ml/h, and 10 ml fractions were collected. The eluate was monitored with a fluorescence detector. The fractions indicated by the horizontal bar were collected and used for further purification.
From: A novel 4-methylumbelliferyl-β-D-xyloside derivative, sulfate-O-3-xylosylβ1-(4-methylumbelliferone), isolated from culture medium of human skin fibroblasts, and its role in methylumbelliferone-initiated glycosaminoglycan biosynthesis
Glycobiology. 1998;8(9): doi: /glycob/
Glycobiology | © 1998 Oxford University Press

3. Fig. 3. Reverse-phase HPLC on Shodex C18-10F
Fig. 3. Reverse-phase HPLC on Shodex C18-10F. HPLC was performed using a Shodex C18-10F column (20 × 250 mm), with a linear gradient of distilled water-acetonitrile. The eluate was monitored with a fluorescence detector. The fractions indicated by the horizontal bar were collected and used for further purification.
From: A novel 4-methylumbelliferyl-β-D-xyloside derivative, sulfate-O-3-xylosylβ1-(4-methylumbelliferone), isolated from culture medium of human skin fibroblasts, and its role in methylumbelliferone-initiated glycosaminoglycan biosynthesis
Glycobiology. 1998;8(9): doi: /glycob/
Glycobiology | © 1998 Oxford University Press

4. Fig. 4. Gel-filtration HPLC on Shodex OHpak KB-802
Fig. 4. Gel-filtration HPLC on Shodex OHpak KB-802. HPLC was performed using a Shodex OHpak KB-802 column (8 × 300 mm) with 20% acetonitrile. The eluate was monitored with a fluorescence detector. The fractions were collected and used for analysis as a purified sample.
From: A novel 4-methylumbelliferyl-β-D-xyloside derivative, sulfate-O-3-xylosylβ1-(4-methylumbelliferone), isolated from culture medium of human skin fibroblasts, and its role in methylumbelliferone-initiated glycosaminoglycan biosynthesis
Glycobiology. 1998;8(9): doi: /glycob/
Glycobiology | © 1998 Oxford University Press

5. Fig. 5. Analysis by HPLC of the Xyl-MU derivative after incubation with various enzymes. (A) before enzymic digestion; (B) after incubation with alkaline phosphatase; (C) after incubation with sulfatase. The arrows denote the positions of Xyl-MU derivatives: 1, novel Xyl-MU derivative; 2, Xyl-MU.
From: A novel 4-methylumbelliferyl-β-D-xyloside derivative, sulfate-O-3-xylosylβ1-(4-methylumbelliferone), isolated from culture medium of human skin fibroblasts, and its role in methylumbelliferone-initiated glycosaminoglycan biosynthesis
Glycobiology. 1998;8(9): doi: /glycob/
Glycobiology | © 1998 Oxford University Press

6. Fig. 6. Mass spectra of the Xyl-MU derivative
Fig. 6. Mass spectra of the Xyl-MU derivative. (A) The Xyl-MU derivative; (B) product ions on tandem mass spectrometric analysis spectrum of the Xyl-MU derivative using m/z as the precursor ion.
From: A novel 4-methylumbelliferyl-β-D-xyloside derivative, sulfate-O-3-xylosylβ1-(4-methylumbelliferone), isolated from culture medium of human skin fibroblasts, and its role in methylumbelliferone-initiated glycosaminoglycan biosynthesis
Glycobiology. 1998;8(9): doi: /glycob/
Glycobiology | © 1998 Oxford University Press

7. Fig. 7. Analysis by HPLC of the Xyl-MU derivative after Smith degradation. (A) and (B) Xyl-MU; (C) and (D), the Xyl-MU derivative. (A) and (C), before Smith degradation; (B) and (D), after Smith degradation.
From: A novel 4-methylumbelliferyl-β-D-xyloside derivative, sulfate-O-3-xylosylβ1-(4-methylumbelliferone), isolated from culture medium of human skin fibroblasts, and its role in methylumbelliferone-initiated glycosaminoglycan biosynthesis
Glycobiology. 1998;8(9): doi: /glycob/
Glycobiology | © 1998 Oxford University Press

8. Fig. 8. Analysis by HPLC of the Xyl-MU derivatives after incubation with [³⁵S]PAPS. Incubation was performed as described in Materials and Methods, using the following acceptors: (A) none; (B) Xyl-MU; (C) Gal-Xyl-MU; (D) Gal-Gal-Xyl-MU.
From: A novel 4-methylumbelliferyl-β-D-xyloside derivative, sulfate-O-3-xylosylβ1-(4-methylumbelliferone), isolated from culture medium of human skin fibroblasts, and its role in methylumbelliferone-initiated glycosaminoglycan biosynthesis
Glycobiology. 1998;8(9): doi: /glycob/
Glycobiology | © 1998 Oxford University Press

9. Fig. 9. Analysis by HPLC of the Xyl-MU derivative after incubation with UDP-[³H]Gal. (A) and (C), before incubation; (B) and (D), after incubation. Incubation was performed as described in Materials and methods, using the following acceptors: (A) and (B), Xyl-MU; (C) and (D), sulfate-O-3-Xyl-MU.
From: A novel 4-methylumbelliferyl-β-D-xyloside derivative, sulfate-O-3-xylosylβ1-(4-methylumbelliferone), isolated from culture medium of human skin fibroblasts, and its role in methylumbelliferone-initiated glycosaminoglycan biosynthesis
Glycobiology. 1998;8(9): doi: /glycob/
Glycobiology | © 1998 Oxford University Press