Reactive oxygen species (ROS) protection via cysteine oxidation in the epidermal cornified cell envelope.

The outermost layer of our skin functions as a barrier to protect us from physical, chemical, and biological environmental insults. This protective function is mediated by the epidermal cornified cell envelope (CE) which serves both as a mechanical and permeability barrier. Recently we have discovered that the CE constitutes also a first-line antioxidant shield which relies greatly on cysteine residues in CE precursor proteins. Here we describe methods and protocols to study the cysteine-mediated antioxidant function of the CE at the level of the whole organ (the skin), individual cells (keratinocytes), or isolated proteins (SPRR family).