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

Authors
  • Sartelet, H
  • Petitfrere, E
  • Martiny, L
  • Haye, B
Type
Published Article
Journal
Biomedical chromatography : BMC
Publication Date
Nov 01, 1999
Volume
13
Issue
7
Pages
465–471
Identifiers
PMID: 10534758
Source
Medline
License
Unknown

Abstract

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.

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