Detection of programmed cell death using fluorescence energy transfer.
Fluorescence energy transfer (FRET) can be generated when green fluorescent protein (GFP) and blue fluorescent protein (BFP) are covalently linked together by a short peptide. Cleavage of this linkage by protease completely eliminates FRET effect. Caspase-3 (CPP32) is an important cellular protease activated during programmed cell death. An 18 amino acid peptide containing CPP32 recognition sequence, DEVD, was used to link GFP and BFP together. CPP32 activation can be monitored by FRET assay during the apoptosis process.
Xu
X
X
Rigel, Inc., 772 Lucerne Drive, Sunnyvale, CA 94086, USA.
Gerard
A L
AL
Huang
B C
BC
Anderson
D C
DC
Payan
D G
DG
Luo
Y
Y
D000595:Amino Acid Sequence; D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases / Q000096:biosynthesis*; D004735:Energy Transfer; D004792:Enzyme Precursors / Q000096:biosynthesis; D005434:Flow Cytometry / Q000379:methods; D015723:Gene Library; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis / Q000096:biosynthesis; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis; D014162:Transfection / Q000379:methods*
D004792:Enzyme Precursors; D008164:Luminescent Proteins; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases