Green fluorescent protein tagging: A novel tool in biomedical research

Abstract

In recent years, the green fluorescent protein (GFP) isolated from the jellyfish, <i style="">Aequorea victoria,</i> has become a novel tool in biomedical research. The tagging of GFP to proteins of interest has widely been used in studies involving various biological events both <i style="">in vitro</i> and <i style="">in vivo</i>. In this report, the authors demonstrate the utility of GFP tagging for studying localization of protein kinase C (PKC<img src='/image/spc_char/alpha.gif'> ), and pleckstrin homology (PH) domains of phospholipase C (PLC<img src='/image/spc_char/delta1.gif'>1), and Akt in mouse neuroblastoma (Neuro 2A) cells. The authors also show the real time translocation of PKC<img src='/image/spc_char/alpha.gif'> and Akt-PH-EGFP to the membrane upon stimulation with either 12-phorbal myristate acetate (PMA) or insulin growth factor (IGF). Pretreatment of cells expressing PKC<img src='/image/spc_char/alpha.gif'> -EGFP or Akt-PH-EGFP with staurosporine (a protein kinase inhibitor) or wortmanin (a phosphatidylinositol 3-kinase inhibitor), prevented the translocation of these proteins to the membrane. These data demonstrate that, GFP tagging could be employed as a tool to study sub cellular localization and distribution of signaling proteins in response to external stimuli in real time.