title: [Fluorescence resonance energy transfer analysis for detecting protein kinase C activation].
abstract: OBJECTIVE
  To establish a sensitive and direct method for detecting the activation of protein kinase C (PKC) using fluorescence resonance energy transfer (FRET) technique.
  
  
  METHODS
  HEK293 cells were transfected with C kinase activity reporter (CKAR) plasmid or/and parathyroid receptor 1 plasmid , and after incubation for 72 h, the fluorescence resonance energy transfer was measured with or without parathyroid or TPA stimulation.
  
  
  RESULTS
  TPA reduced the efficiency of FRET and increased the emission ratio of CFP/YFP (C/Y) in HEK293 cells transfected with CKAR. PTH(1-34) could increase the emission ratio of C/Y in HEK293 cells co-transfected with CKAR and PTHR1 but not in cells transfected with CKAR.
  
  
  CONCLUSIONS
  FRET analysis using CKAR can be utilized to detect the activation of PKC, which provides a useful means for studying the signaling pathways associated with PKC.
authors: Department of Spinal Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. ljqiskobebryant@163.com
authors: Jun-qing
authors: JQ
authors: Li
authors: Jian-ming
authors: JM
authors: Jiang
authors: Rui-qiang
authors: RQ
authors: Feng
authors: Hui
authors: H
authors: Jiang
authors: Wen-juan
authors: WJ
authors: Yan
authors: De-hong
authors: DH
authors: Yang
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D004789:Enzyme Activation; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry; D011493:Protein Kinase C / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D011493:Protein Kinase C