Cell migration plays an important role in cancer metastasis and wound healing. It is well known that reactive oxygen species (ROS) can modulate cell migration in a variety of cell types. ROS play a vital role in redox homeostasis via a variety of post-translational modifications. S-glutathionylation is one of the major ROS-mediated oxidative protein modifications that regulate various biological processes such as cell migration, invasion, and apoptosis. However, underlying mechanisms of how S-glutathionylation regulates signaling pathways associated with cell migration are not completely understood due to limited information on redox regulatory proteins involved in migration. With the use of Cytoscape and DAVID GO tools, we have narrowed down thirty-eight redox regulatory proteins and their specific cysteine that are related to cell migration under physiological stress.<br/><br/>In this work, we have validated nine migration-related proteins out of 38 through wound healing assays. The assay was conducted with wild-type and specific cysteine to serine mutants of the candidates under 25 mM and 5 mM glucose conditions. Interestingly, three of them (NISCH, PP2CA, ARHGEF7) resulted in an increase in migration under low glucose conditions upon glutathionylation of specific cysteines. Further studies on PP2Cα, one of three proteins indicated that PP2CA C314 glutathionylation increases cell migration via JNK and ERK signaling pathways. These identifications of specific redox-regulated cysteines may lead to understanding redox-mediated mechanisms underlying cell migration. Moreover, we observed that MCF7 cells increase migration in a D-alanine dose-dependent manner when DAAO (D amino acid oxidase) is overexpressed in the cytoplasm. This allows us to produce hydrogen peroxide in a spatial and controllable manner mimicking the physiological conditions leading to the discovery of novel cysteines associated with migration through proteomic approaches. These remarkable findings may lead to identifying more redox-regulated cysteines modulating migration.

Identification of glutathionylated cysteines regulating cell migration

ACS Spring 2022

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