title: Peroxynitrite-dependent upregulation of SRC kinases in red blood cells: strategies to study the activation mechanisms.
abstract: Several studies have demonstrated that treatment of cells with oxidants, and in particular with peroxynitrite, may cause the upregulation of tyrosine phosphorylation signaling. In erythrocytes, peroxynitrite induces tyrosine phosphorylation of the major intrinsic membrane protein, band 3. A closer look at the enzymes involved revealed that the effect of peroxynitrite was due to the inhibition of phosphotyrosine phosphatases and/or to the activation of src kinases. The activity of src kinases is modulated not only by phosphatases and other kinases but also through redox modification of cysteine residues: Peroxynitrite can, thus, affect src kinase activity by means of direct and indirect mechanisms. In this chapter, we describe the different pathways leading to src kinase activation and the experimental procedures that can be performed to reveal the activation mechanism. The aim is to provide a more general strategy adaptable to different cell types and different oxidants.
authors.affiliation: Antioxidant Research Laboratory, Unit of Human Nutrition, National Institute for Food and Nutrition Research, Rome, Italy.
authors.forename: Mauro
authors.initials: M
authors.lastname: Serafini
authors.forename: Cinzia
authors.initials: C
authors.lastname: Mallozzi
authors.forename: Anna Maria Michela
authors.initials: AM
authors.lastname: Di Stasi
authors.forename: Maurizio
authors.initials: M
authors.lastname: Minetti
mesh_terms: D000595:Amino Acid Sequence; D004789:Enzyme Activation; D004912:Erythrocytes / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D030421:Peroxynitrous Acid / Q000494:pharmacology*; D010766:Phosphorylation; D015854:Up-Regulation; D019061:src-Family Kinases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D030421:Peroxynitrous Acid; D019061:src-Family Kinases