Paper Citing NAMD - Abstract
Chen, Kun-Ming; Spratt, Thomas E.; Stanley, Bruce A.; De Cotiis, Dan A.; Bewley, Maria C.; Flanagan, John M.; Desai, Dhimant; Das, Arunangshu; Fiala, Emerich S.; Amin, Shantu; El-Bayoumy, Karam
Inhibition of nuclear Factor-kappa B DNA binding by organoselenocyanates through covalent modification of the p50 subunit
CANCER RESEARCH, 67:10475-10483, NOV 1 2007
Most known chemopreventive agents including certain selenium compounds suppress the activation of the nuclear factor kappa B (NF-kappa B), but the mechanisms remain largely elusive. Toward this end, we initially showed that the inhibition of NF-kappa B DNA binding by benzyl selenocyanate (BSC) and 1,4-phenylenebis(methylene)selenocyanate (p-XSC) was reversed by the addition of DTT; this suggests the formation of DTT-reducible selenium-sulfur bonds between selenocyanate moieties and cysteine residues in NF-kappa B (p50) protein. Furthermore, the inhibitory effect of selenocyanates on NF-kappa B was not altered in the presence of physiologic level of reduced glutathione (1 mmol/L), suggesting that selenocyanates can also inhibit NF-kappa B in vivo. Using both matrix-assisted laser desorption/ionization-time of flight and tandem mass spectrometry fragmentation, we showed for the first time that the Cys(62) residue in the active site of NF-kappa B (p50) protein was modified by BSC through the formation of a selenium-sulfur bond. In addition, p-XSC-bound NF-kappa B (p50) protein was also detected by a radiotracer method. To provide further support, molecular models of both BSC and p-XSC positioned in the DNA binding pocket of the p50 were constructed through the covalent modification of Cys(62); the models reveal that DNA substrate could be hindered to enter its DNA binding region. This study shows for the first time that BSC and p-XSC may exert their chemopreventive activity, at least in part, by inhibiting NF-kappa B through covalent modification of Cys(62) of the p50 subunit of NF-kappa B.
