Screening assay for the identification of deoxyhypusine synthase inhibitors.
The 1st step in the posttranslational hypusine [N(epsilon)-(4-amino-2-hydroxybutyl)lysine] modification of eukaryotic translation initiation factor 5A (eIF5A) is catalyzed by deoxyhypusine synthase (DHS). The eIF5A intermediate is subsequently hydroxylated by deoxyhypusine hydroxylase (DHH), thereby converting the eIF5A precursor into a biologically active protein. Depletion of eIF5A causes inhibition of cell growth, and the identification of eIF5A as a cofactor of the HIV Rev protein turns this host protein and therefore DHS into an interesting target for drugs against abnormal cell growth and/or HIV replication. The authors developed a 96-well format DHS assay applicable for the screening of DHS inhibitors. Using this assay, they demonstrate DHS inhibition by AXD455 (Semapimod, CNI-1493). This assay represents a powerful tool for the identification of new DHS inhibitors with potency against cancer and HIV.
Department of Assay Development and Screening, Axxima Pharmaceuticals AG, Max-Lebsche-Platz 32, D-81377 München, Germany. marc.sommer@axxima.com
Marc-Nicola
MN
Sommer
Dorian
D
Bevec
Bert
B
Klebl
Birgit
B
Flicke
Kerstin
K
Hölscher
Tatjana
T
Freudenreich
Ilona
I
Hauber
Joachim
J
Hauber
Helmut
H
Mett
D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D006835:Hydrazones / Q000737:chemistry* / Q000494:pharmacology; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D010448:Peptide Initiation Factors / Q000302:isolation & purification; D016601:RNA-Binding Proteins / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification; D000097574:Eukaryotic Translation Initiation Factor 5A; D000813:Anilides; D006146:Guanidines
D004791:Enzyme Inhibitors; D006835:Hydrazones; D000587:Oxidoreductases Acting on CH-NH Group Donors; D010448:Peptide Initiation Factors; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D000097574:Eukaryotic Translation Initiation Factor 5A; C097256:semapimod; D000813:Anilides; D006146:Guanidines; C063507:deoxyhypusine synthase