The use of green fluorescent proteins to view association between phospholipase C beta and G protein subunits in cells.
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.
D000818:Animals; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D044385:GTP-Binding Protein alpha Subunits / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics*; D008967:Molecular Biology / Q000379:methods; D016716:PC12 Cells; D054799:Phospholipase C beta; D051381:Rats; D015398:Signal Transduction; D010738:Type C Phospholipases / Q000235:genetics* / Q000378:metabolism
D044385:GTP-Binding Protein alpha Subunits; D007202:Indicators and Reagents; D007527:Isoenzymes; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D010738:Type C Phospholipases; D054799:Phospholipase C beta