A new assay based on fluorescence resonance energy transfer to determine the binding affinity of Bcl-xL inhibitors.
We developed a new assay of Bcl-xL inhibitors based on fluorescence resonance energy transfer that occurs between an AEDANS-labeled Bak-BH3 peptide and three tryptophans in the BH1 and BH2 domains of Bcl-xL. The method can tolerate up to 5% DMSO, and it was validated with several Bcl-xL inhibitors. It can be adapted to screen for compounds targeting other Bcl-2 family proteins.
Feng
Yu
Y
Department of Molecular Pharmacology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.
Shen
Xu
X
Chen
Kaixian
K
Jiang
Hualiang
H
Liu
Dongxiang
D
D017209:Apoptosis; D001665:Binding Sites; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes; D015336:Molecular Probe Techniques; D010455:Peptides; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014364:Tryptophan; D051020:bcl-X Protein / Q000037:antagonists & inhibitors*
D005456:Fluorescent Dyes; D010455:Peptides; D051020:bcl-X Protein; D014364:Tryptophan