Purification and analysis of the neutral glycan moiety of glycosyl phosphatidylinositol from porcine thyroid cells

✍ Scribed by Herve Sartelet; Emmanuelle Petitfrere; Laurent Martiny; Bernard Haye

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 161 KB
• Volume: 13
• Category: Article
• ISSN: 0269-3879
• DOI: 10.1002/(sici)1099-0801(199911)13:7<465::aid-bmc913>3.0.co;2-d

✦ Synopsis

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.