Purification and analysis of the neutral glycan moiety of glycosyl phosphatidylinositol from porcine thyroid cells.
Metabolic labelling of inositolphosphate glycan with radioactive precursors is not sufficient to characterize and assess the involvement of the glycosyl phosphatidylinositol/inositolphosphate glycan (GPI/IPG) system in porcine thyroid cell signal transduction machinery. A protocol is described for the isolation and purification of free GPI using differential polarity of lipids and sequential thin layer chromatography. The purification until homogeneity of GPI constitutes a required step for gas chromatographic analysis. Next, successive chemical treatments allowed us to remove the neutral glycan moiety of thyroidal GPI, and its composition was obtained by gas chromatography. The proposed structure is consistent with data available for GPI anchor, but differs from compositional analysis data reported for insulin-sensitive GPI. Our results support the existence in porcine thyroid cells of the GPI/IPG system, which can take part in TSH-dependent signal transduction processes.
Sartelet
H
H
Université de Reims-Champagne Ardenne, IFR 53 Biomolécules, UPRES-A CNRS 6021, Laboratoire de Biochimie, UFR Sciences de Reims, Moulin de la housse, BP 1039, 51687 Reims Cedex 2, France.
Petitfrere
E
E
Martiny
L
L
Haye
B
B
D000818:Animals; D002478:Cells, Cultured; D002849:Chromatography, Gas / Q000379:methods*; D002855:Chromatography, Thin Layer / Q000379:methods*; D017261:Glycosylphosphatidylinositols / Q000737:chemistry*; D011134:Polysaccharides / Q000302:isolation & purification*; D011849:Radioactive Tracers; D013552:Swine; D013961:Thyroid Gland / Q000737:chemistry* / Q000166:cytology
D017261:Glycosylphosphatidylinositols; D011134:Polysaccharides; D011849:Radioactive Tracers