Identification of potential cellular targets of aloisine A by affinity chromatography.
Affinity chromatography was used to identify potential cellular targets of aloisine A (7-n-butyl-6-(4'-hydroxyphenyl)-5H-pyrrolo[2,3b]pyrazine), a potent inhibitor of cyclin-dependent kinases. This technique is based on the immobilization of the drug on a solid matrix, followed by identification of specifically bound proteins. To this end, both aloisine A and the protein-kinase inactive control N-methyl aloisine, bearing extended linker chains have been synthesized. We present the preparation of such analogues having the triethylene glycol chain at different positions of the molecule, as well as their immobilization on an agarose-based matrix. Affinity chromatography of various biological extracts on the aloisine matrices allowed the identification of both protein kinases and non-kinase proteins as potential cellular targets of aloisine.
Corbel
Caroline
C
CNRS USR-3151, Protein Phosphorylation and Human Disease, Station Biologique, B.P.74, F-29682 Roscoff Cedex, Bretagne, France.
Haddoub
Rose
R
Guiffant
Damien
D
Lozach
Olivier
O
Gueyrard
David
D
Lemoine
Jérôme
J
Ratin
Morgane
M
Meijer
Laurent
L
Bach
Stéphane
S
Goekjian
Peter
P
D000818:Animals; D001921:Brain / Q000201:enzymology; D002846:Chromatography, Affinity / Q000379:methods*; D018844:Cyclin-Dependent Kinases / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism; D010963:Plasmodium falciparum / Q000201:enzymology; D011485:Protein Binding; D011719:Pyrazines / Q000737:chemistry* / Q000494:pharmacology*; D011758:Pyrroles / Q000737:chemistry* / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013552:Swine
C544033:7-n-butyl-6-(4'-hydroxyphenyl)-5H-pyrrolo(2,3b)pyrazine; D011719:Pyrazines; D011758:Pyrroles; D018844:Cyclin-Dependent Kinases; D038362:Glycogen Synthase Kinase 3