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Glycosylinositol phosphorylceramides from Rosa cell cultures are boron-bridged in the plasma membrane and form complexes with rhamnogalacturonan II.

Voxeur A, Fry SC - Plant J. (2014)

Bottom Line: Using high-voltage paper electrophoresis, we showed that addition of GIPCs decreased the electrophoretic mobility of radiolabelled RG-II, suggesting formation of a GIPC-B-RG-II complex.We conclude that B plays a structural role in the plasma membrane.Finally, our results suggest a role for GIPCs in the RG-II dimerization process.

View Article: PubMed Central - PubMed

Affiliation: The Edinburgh Cell Wall Group, Institute of Molecular Plant Sciences, The University of Edinburgh, Edinburgh, EH9 3JH, UK.

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Purification of glycosylinositol phosphorylceramide (GIPC) from Rosa cell culture. (a) GIPC purification scheme adapted from Buré et al. (2011). (b) Thin-layer chromatography (TLC) of the phase A, B and C obtained during the step shaded grey in (a). Suc, sucrose marker.
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fig03: Purification of glycosylinositol phosphorylceramide (GIPC) from Rosa cell culture. (a) GIPC purification scheme adapted from Buré et al. (2011). (b) Thin-layer chromatography (TLC) of the phase A, B and C obtained during the step shaded grey in (a). Suc, sucrose marker.

Mentions: In order to test for possible RG-II–GIPC interactions, we tried to obtain GIPC as pure as possible by adapting the method of Buré et al. (2011). The main difference from the original protocol was the addition of a phase-partition step with methanol/chloroform/formic acid/water (Figure3a). As judged by TLC of the two phases obtained, the less polar lipids were found in the organic phase, the contaminating sugars in the AP and the GIPC at the interface (Figure3b). This interface material was then submitted to an acidic butanol/water phase partition, which removed polymers. The purified GIPC-containing BP was collected.


Glycosylinositol phosphorylceramides from Rosa cell cultures are boron-bridged in the plasma membrane and form complexes with rhamnogalacturonan II.

Voxeur A, Fry SC - Plant J. (2014)

Purification of glycosylinositol phosphorylceramide (GIPC) from Rosa cell culture. (a) GIPC purification scheme adapted from Buré et al. (2011). (b) Thin-layer chromatography (TLC) of the phase A, B and C obtained during the step shaded grey in (a). Suc, sucrose marker.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4230332&req=5

fig03: Purification of glycosylinositol phosphorylceramide (GIPC) from Rosa cell culture. (a) GIPC purification scheme adapted from Buré et al. (2011). (b) Thin-layer chromatography (TLC) of the phase A, B and C obtained during the step shaded grey in (a). Suc, sucrose marker.
Mentions: In order to test for possible RG-II–GIPC interactions, we tried to obtain GIPC as pure as possible by adapting the method of Buré et al. (2011). The main difference from the original protocol was the addition of a phase-partition step with methanol/chloroform/formic acid/water (Figure3a). As judged by TLC of the two phases obtained, the less polar lipids were found in the organic phase, the contaminating sugars in the AP and the GIPC at the interface (Figure3b). This interface material was then submitted to an acidic butanol/water phase partition, which removed polymers. The purified GIPC-containing BP was collected.

Bottom Line: Using high-voltage paper electrophoresis, we showed that addition of GIPCs decreased the electrophoretic mobility of radiolabelled RG-II, suggesting formation of a GIPC-B-RG-II complex.We conclude that B plays a structural role in the plasma membrane.Finally, our results suggest a role for GIPCs in the RG-II dimerization process.

View Article: PubMed Central - PubMed

Affiliation: The Edinburgh Cell Wall Group, Institute of Molecular Plant Sciences, The University of Edinburgh, Edinburgh, EH9 3JH, UK.

Show MeSH
Related in: MedlinePlus