Analysis of inositol phospholipid turnover during lymphocyte activation.
Receptor-mediated activation of phospholipase C (PLC) leads to the hydrolysis of membrane inositol phospholipids, generating diacylglycerol (DAG) and water-soluble inositol phosphates. This signaling mechanism is used by antigen receptors on T and B cells that have been implicated as mediators of receptor-induced influx of extracellular Ca(2+). This unit provides protocols that describe the resolution of InsP by Dowex anion-exchange chromatography. This technique provides a reliable means of separating inositol monophosphate, inositol bisphosphate, and inositol trisphosphate, but does not resolve isomers of these. An Alternate Protocol describes the separation of inositol phosphates by anion-exchange HPLC. A protocol for resolution of inositol phospholipids by thin-layer chromatography (TLC) is also provided.
Imboden
J B
JB
University of California and Veterans Affairs Medical Center, San Francisco, California, USA.
Shoback
D M
DM
Inokuchi
S
S
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D002855:Chromatography, Thin Layer / Q000379:methods; D006801:Humans; D006868:Hydrolysis; D007295:Inositol Phosphates / Q000032:analysis; D008213:Lymphocyte Activation / Q000276:immunology*; D008214:Lymphocytes / Q000737:chemistry / Q000276:immunology*; D010716:Phosphatidylinositols / Q000032:analysis / Q000378:metabolism*; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D010738:Type C Phospholipases / Q000737:chemistry
D007295:Inositol Phosphates; D010716:Phosphatidylinositols; D011948:Receptors, Antigen, T-Cell; D010738:Type C Phospholipases