Reactive oxygen species (ROS) and especially hydrogen peroxide, are potent signaling molecules that activate cellular defense responses. Hydrogen peroxide can provoke reversible and irreversible oxidative posttranslational modifications on cysteine residues of proteins that act in diverse signaling circuits. The initial oxidation product of cysteine, sulfenic acid, has emerged as a biologically relevant posttranslational modification, because it is the primary sulfur oxygen modification that precedes divergent series of additional adaptations. In this review, we focus on the functional consequences of sulfenylation for both mammalian and plant proteins. Furthermore, we created compendia of sulfenylated proteins in human and plants based on mass spectrometry experiments, thereby defining the current plant and human sulfenomes. To assess the evolutionary conservation of sulfenylation, the sulfenomes of human and plants were compared based on protein homology. In total, 185 human sulfenylated proteins showed homology to sulfenylated plant proteins and the conserved sulfenylation targets participated in specific biological pathways and metabolic processes. Comprehensive functional studies of sulfenylation remains a future challenge, with multiple candidates suggested by mass spectrometry awaiting scrutinization.