title: Nitration of tyrosines 46 and 48 induces the specific degradation of cytochrome c upon change of the heme iron state to high-spin.
abstract: The Reactive Nitrogen and Oxygen Species (the so-called RNOS), which are well-known radicals formed in the mitochondria under nitro-oxidative cell stress, are responsible for nitration of tyrosines in a wide variety of proteins and, in particular, in cytochrome c (Cc). Only three out of the five tyrosine residues of human Cc, namely those at positions 67, 74 and 97, have been detected in vivo as nitrotyrosines. However, nitration of the two other tyrosines, namely those at positions 46 and 48, has never been detected in vivo despite they are both well-exposed to solvent. Here we investigate the changes in heme coordination and alkaline transition, along with the peroxidase activity and in cell degradation of Cc mutants in which all their tyrosine residues - with the only exception of that at position 46 or 48 - are replaced by phenylalanines. In Jurkat cell extracts devoid of proteases inhibitors, only the high-spin iron nitrated forms of these monotyrosine mutants are degraded. Altogether the resulting data suggest that nitration of tyrosines 46 and 48 makes Cc easily degradable upon turning the heme iron state to high-spin.
authors.affiliation: Instituto de Bioquímica Vegetal y Fotosíntesis, cicCartuja, Universidad de Sevilla-CSIC, Americo Vespucio 49, Sevilla 41092, Spain.
authors.forename: Irene
authors.initials: I
authors.lastname: Díaz-Moreno
authors.forename: José M
authors.initials: JM
authors.lastname: García-Heredia
authors.forename: Antonio
authors.initials: A
authors.lastname: Díaz-Quintana
authors.forename: Miguel
authors.initials: M
authors.lastname: Teixeira
authors.forename: Miguel A
authors.initials: MA
authors.lastname: De la Rosa
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics; D045304:Cytochromes c / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006418:Heme / Q000737:chemistry*; D006736:Horses; D006801:Humans; D007501:Iron / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009566:Nitrates / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D017433:Protein Structure, Secondary; D026361:Reactive Nitrogen Species / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D016415:Sequence Alignment; D014443:Tyrosine / Q000737:chemistry / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; D009566:Nitrates; D026361:Reactive Nitrogen Species; D017382:Reactive Oxygen Species; D014443:Tyrosine; D006418:Heme; D045304:Cytochromes c; D007501:Iron