Identification of actin as quercetin-binding protein: an approach to identify target molecules for specific ligands.
The biological effect of flavonoids is commonly studied by assaying the activity of a protein of interest. Taking a reverse approach, we identified target proteins of the widely studied flavonol quercetin by exploiting the altered spectroscopic properties of target proteins and ligands on their molecular interaction. Nuclear extracts of human leukemia cells were fractionated by column chromatography and assayed for their ability to alter the fluorescence emission spectra, and finally the proteins present in fractions of interest were identified by mass spectrometry. Among the identified proteins, actin was shown to be a quercetin-binding nuclear protein.
Böhl
M
M
Institute of Zoology, University of Technology Dresden, D-01062 Dresden, Germany.
Czupalla
C
C
Tokalov
S V
SV
Hoflack
B
B
Gutzeit
H O
HO
D000199:Actins / Q000378:metabolism*; D002458:Cell Fractionation; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D018922:HL-60 Cells / Q000737:chemistry; D006801:Humans; D008024:Ligands; D009687:Nuclear Proteins / Q000302:isolation & purification; D011485:Protein Binding; D011794:Quercetin / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000199:Actins; D008024:Ligands; D009687:Nuclear Proteins; D011794:Quercetin