Protocol

Inositol Phosphates and Lipids

Volume 645 of the series Methods in Molecular Biology pp 47-60

Date:

Molecular Manipulation and Analysis of Inositol Phosphate and Pyrophosphate Levels in Mammalian Cells

  • James C. OttoAffiliated withDepartment of Pharmacology and Cancer Biology, Howard Hughes Medical Institute, Duke University Medical Center
  • , John D. YorkAffiliated withDepartment of Pharmacology and Cancer Biology, Howard Hughes Medical Institute, Duke University Medical Center

* Final gross prices may vary according to local VAT.

Get Access

Abstract

Lipid-derived inositol phosphates (InsPs) comprise a family of second messengers that arise through the action of six classes of InsP kinases, generally referred to as IPKs. Genetic studies have indicated that InsPs play critical roles in embryonic development, but the mechanisms of action for InsPs in mammalian cellular function are largely unknown. This chapter outlines a method for manipulating cellular InsP profiles through the coexpression of a constitutively active G protein and various IPKs. It provides a mechanism by which the metabolism of a variety of InsPs can be upregulated, enabling the evaluation of the effects of these InsPs on cellular functions.

Key words:

Inositol phosphate G protein Phospholipase C HPLC Second messenger Signal transduction