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The Use of Green Fluorescent Proteins to View Association Between Phospholipase Cβ and G Protein Subunits in Cells

  • Suzanne Scarlata
  • Louisa Dowal
Part of the Methods in Molecular Biology™ book series (MIMB, volume 237)

Summary

A major advance in biology is the ability to attach either green fluorescence protein (GFP) or one of its variants to a target protein and follow its cellular localization and interaction with other partners by fluorescence microscopy. Our laboratory has previously developed fluorescence energy-transfer methods to measure the kinetics and affinities of the lateral association between phospholipase C (PLC) and G protein subunits on membrane surfaces. We are currently developing methods to view these associations in living cells using fluorescence resonance energy transfer (FRET) between GFP-based chimeras. Because the improvements and variations of these GFP-based FRET techniques has continued on a rapid pace, we focus only on the basic principles behind these measurements and the methods used, which may continue to be applicable as improvements become available.

Key Words

Phospholipase Cβ G proteins fluorescence resonance energy transfer fluorescence microscopy green fluorescent protein cell culture 

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Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Suzanne Scarlata
    • 1
  • Louisa Dowal
    • 1
  1. 1.Department of Physiology and BiophysicsState University of New York at Stony BrookStony Brook

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