Design and characterization of an active site selective caspase-3 transnitrosating agent.
The oxidative addition of nitric oxide (NO) to a thiol, S-nitrosation, is a focus of studies on cyclic guanosine monophosphate (cGMP)-independent NO signaling. S-Nitrosation of the catalytic cysteine of the caspase proteases has important effects on apoptosis and consequently has received attention. Here we report on a small molecule that can directly probe the effects of S-nitrosation on the caspase cascade. This chemical tool is capable of permeating the mammalian cell membrane, selectively transnitrosating the caspase-3 active site cysteine, and halting apoptosis in cultured human T-cells. The efficacy of this reagent was compared with the commonly used reagent S-nitrosoglutathione and an esterified derivative.
Mitchell
Douglas A
DA
Department of Chemistry, University of California, Berkeley, California 94720-1460, USA.
Morton
Sarah U
SU
Marletta
Michael A
MA
D017209:Apoptosis; D001665:Binding Sites; D001710:Biotin / Q000737:chemistry; D053148:Caspase 3 / Q000737:chemistry*; D053453:Caspase 9 / Q000378:metabolism; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D006152:Cyclic GMP / Q000378:metabolism; D003545:Cysteine / Q000737:chemistry; D015195:Drug Design; D004789:Enzyme Activation; D006801:Humans; D007700:Kinetics; D009569:Nitric Oxide / Q000737:chemistry* / Q000378:metabolism; D015538:Nitrosation; D010455:Peptides / Q000737:chemistry
D010455:Peptides; D009569:Nitric Oxide; D001710:Biotin; D053148:Caspase 3; D053453:Caspase 9; D006152:Cyclic GMP; D003545:Cysteine