Different phosphorylated forms of inositolphosphate glycan could be involved in the transforming growth factor-beta 1 (TGF-beta 1) signalling pathway.
Labelling with [3H]glucosamine was used to prepare a transforming growth factor-beta 1 (TGF-beta 1)-sensitive glycosylphosphatidylinositol (GPI) from monolayer cultures of rabbit articular chondrocytes (RAC), which may be involved in control of the cell cycle. The polar headgroup of this glycosylphosphatidylinositol was generated by both phosphatidylinositol-specific phospholipase C (PI-PLC) and pronase E digestion. The molecule emerged in only one peak on a Dowex AG1-X8 chromatogram, eluted at 0.1 N ammonium formate. In contrast, similar experiments performed on cellular extract from cultures previously labelled with [3H]glucosamine displayed four radioactive peaks eluting at 0.1, 0.2, 0.5 and 1 N ammonium formate, respectively. Evidence that the eluting position of these peaks was dependent on the number of phosphate residues present in each fraction was demonstrated by both [32P]phosphorus labelling and change in the position of alkaline phosphatase-induced shift in elution volume. We also demonstrated that the GPI-derived inositolphosphate glycan (IPG) could be hyperphosphorylated into the cell under the action of a kinase whose activity was enhanced by TGF-beta 1 itself. We have also shown that all of these IPG forms could mimic the TGF-beta-induced increase of DNA replication rate of RAC, with a higher activity for peaks III and IV than peaks I and II.
Vivien
D
D
Laboratoire de Biochimie du Tissu Conjonctif, CJF INSERM 91-06, CHU Caen Côte de Nacre, France.
Bogdanowicz
P
P
Boumediene
K
K
Martiny
L
L
Haye
B
B
Pujol
J P
JP
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology / Q000378:metabolism; D002453:Cell Cycle; D002455:Cell Division; D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange; D004261:DNA Replication / Q000187:drug effects; D005944:Glucosamine / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism*; D010766:Phosphorylation; D011134:Polysaccharides / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism; D011817:Rabbits; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000378:metabolism*
D017261:Glycosylphosphatidylinositols; D007295:Inositol Phosphates; D011134:Polysaccharides; D016212:Transforming Growth Factor beta; C028039:inositol phosphate glycan; D011494:Protein Kinases; D005944:Glucosamine