Detection of nitrosothiols and other nitroso species in vitro and in cells

Abstract

The nitric oxide (NO)-mediated nitrosation of peptides and proteins may play important roles in normobiology and pathobiology. With the realization that S-nitrosothiols (RSNOs) participate in the transport, storage, and delivery of NO, as well as posttranslational modifications in cell signaling and inflammatory processes, there is an increasing need for the detection of nitrosothiols (RSNOs) and other nitroso species in cells and tissues. In this chapter, we describe the utilization of a gas phase chemiluminescence-based assay and "biotin switch" method for the detection of nitroso species in cells. These methods are sensitive enough to quantify and contrast the different pools of nitroso species that may coexist under physiologically relevant conditions. They also provide the means to characterize and identify proteins that may represent specific targets for nitrosation reactions.