The NADPH oxidase (NOX) family of proteins is involved in regulating many diverse
cellular processes, which is largely mediated by NOX-mediated reversible oxidation
of target proteins in a process known as redox signaling. Protein cysteine residues
are the most prominent targets in redox signaling, and to understand the mechanisms
by which NOX affect cellular pathways, specific methodology is required to detect
specific oxidative cysteine modifications and to identify targeted proteins. Among
the many potential redox modifications involving cysteine residues, reversible modifications
most relevant to NOX are sulfenylation (P-SOH) and S-glutathionylation (P-SSG), as
both can induce structural or functional alterations. Various experimental approaches
have been developed to detect these specific modifications, and this chapter will
detail state-of-the-art methodology to selectively evaluate these modifications in
specific target proteins in relation to NOX activation. We also discuss some of the
limitations of these procedures and potential complementary approaches.
