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Viktoria Petrova authoredViktoria Petrova authored
- Figure 3 B) caption
- Figure 3 B) source
- Figure 4 A) caption
- Figure 4 A) source
- Figure 4 B) caption
- Figure 4 B) source
- Figure 4 C) caption
- Figure 4 C) source
- Supplementary Figure 4 A) caption
- Supplementary Figure 4 A) source
- Supplementary Figure 4 B) caption
- Supplementary Figure 4 B) source
- Supplementary Figure 5 A) caption
- Supplementary Figure 5 A) source
Figure 3 B) caption
Figure 3 B). Analysis of the β-1,3-glycosidic activity of heterologously expressed and purified GBP1 and mutant GBP1E500A on laminarin from Laminaria digitata. After overnight incubation at 25°C, the digestion products were analyzed by thin-layer chromatography. The experiment was repeated at least four times with similar results.
Figure 3 B) source
Figure 4 A) caption
Figure 4 A). The digestion products resulting from the enzymatic breakdown of laminarin by GBP1 over time were analyzed using thin-layer chromatography (TLC). Untreated (UT) and GBP1E500A-treated laminarin (20 h) served as controls. Digestion was performed at 25°C.
Figure 4 A) source
Figure 4 B) caption
Figure 4 B). Quantification of GBP1-catalyzed laminarin digestion products. TLC band intensity was quantified using the ImageJ software.
Figure 4 B) source
Figure 4 C) caption
Figure 4 C). Laminarin (4 mg/mL-1) was digested with GBP1 (70 nM) for 10 min at different temperatures (left graph) or in different buffers (10 mM, pH 5–9, right graph). Products from GBP1-catalyzed laminarin digestion assays were separated by TLC (Figures S4A and S4B) and quantified via ImageJ. UT, untreated.
Figure 4 C) source
Supplementary Figure 4 A) caption
Supplementary Figure 4 A). Laminarin (4 mg·mL−1) was digested with GBP1 (70 nM) for 10 min at different temperatures. Sample without enzymes (UT) was mock-digested at 80 °C. Products from GBP1-catalyzed laminarin digestion assays in (A) and (B) were separated by thin layer chromatography. Quantification of thin layer chromatographs in Figure 4C.
Supplementary Figure 4 A) source
https://www.cell.com/current-biology/pdfExtended/S0960-9822(23)01449-5
Supplementary Figure 4 B) caption
Supplementary Figure 4 B). Laminarin (4 mg·mL−1) was digested with purified GBP1 (70 nM) in different buffers (10 mM, pH 5-9). Digestions were performed at 60 °C for 10 min. Products from GBP1- catalyzed laminarin digestion assays in (A) and (B) were separated by thin layer chromatography. Quantification of thin layer chromatographs in Figure 4C.
Supplementary Figure 4 B) source
https://www.cell.com/current-biology/pdfExtended/S0960-9822(23)01449-5
Supplementary Figure 5 A) caption
Supplementary Figure 5 A). Laminarin from E. bicyclis (2.4 mM) was digested with GBP1 (72 nM) for 1 h at 60 °C. The digestion products were analyzed by thin layer chromatography.
Supplementary Figure 5 A) source
https://www.cell.com/current-biology/pdfExtended/S0960-9822(23)01449-5