title: Structural basis of mitochondrial dysfunction in response to cytochromecphosphorylation at tyrosine 48.
abstract: Regulation of mitochondrial activity allows cells to adapt to changing conditions and to control oxidative stress, and its dysfunction can lead to hypoxia-dependent pathologies such as ischemia and cancer. Although cytochromecphosphorylation-in particular, at tyrosine 48-is a key modulator of mitochondrial signaling, its action and molecular basis remain unknown. Here we mimic phosphorylation of cytochromecby replacing tyrosine 48 withp-carboxy-methyl-l-phenylalanine (pCMF). The NMR structure of the resulting mutant reveals significant conformational shifts and enhanced dynamics aroundpCMF that could explain changes observed in its functionality: The phosphomimetic mutation impairs cytochromecdiffusion between respiratory complexes, enhances hemeprotein peroxidase and reactive oxygen species scavenging activities, and hinders caspase-dependent apoptosis. Our findings provide a framework to further investigate the modulation of mitochondrial activity by phosphorylated cytochromecand to develop novel therapeutic approaches based on its prosurvival effects.
authors.affiliation: Instituto de Investigaciones Químicas, cicCartuja, Universidad de Sevilla-Spanish National Scientific Council (CSIC), 41092 Seville, Spain.
authors.forename: Blas
authors.initials: B
authors.lastname: Moreno-Beltrán
authors.affiliation: Instituto de Investigaciones Químicas, cicCartuja, Universidad de Sevilla-Spanish National Scientific Council (CSIC), 41092 Seville, Spain.
authors.forename: Alejandra
authors.initials: A
authors.lastname: Guerra-Castellano
authors.affiliation: Instituto de Investigaciones Químicas, cicCartuja, Universidad de Sevilla-Spanish National Scientific Council (CSIC), 41092 Seville, Spain.
authors.forename: Antonio
authors.identifier: 0000-0001-8973-8009
authors.initials: A
authors.lastname: Díaz-Quintana
authors.affiliation: Magnetic Resonance Center (CERM), Department of Chemistry, University of Florence, 50019 Sesto Fiorentino, Florence, Italy.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Del Conte
authors.affiliation: Instituto de Investigaciones Químicas, cicCartuja, Universidad de Sevilla-Spanish National Scientific Council (CSIC), 41092 Seville, Spain.
authors.forename: Sofía M
authors.initials: SM
authors.lastname: García-Mauriño
authors.affiliation: Diamond Light Source, Didcot, Oxfordshire OX11 0DE, United Kingdom.
authors.forename: Sofía
authors.initials: S
authors.lastname: Díaz-Moreno
authors.affiliation: Instituto de Investigaciones Químicas, cicCartuja, Universidad de Sevilla-Spanish National Scientific Council (CSIC), 41092 Seville, Spain.
authors.forename: Katiuska
authors.initials: K
authors.lastname: González-Arzola
authors.affiliation: Centro Andaluz de Biología del Desarrollo, Universidad Pablo de Olavide-CSIC, 41013 Seville, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Santos-Ocaña
authors.affiliation: Institute of Biocomputation and Physics of Complex Systems (BIFI), Joint Unit BIFI-Instituto Química-Física Rocasolano (CSIC), Universidad de Zaragoza, 50018 Zaragoza, Spain.
authors.forename: Adrián
authors.initials: A
authors.lastname: Velázquez-Campoy
authors.affiliation: Instituto de Investigaciones Químicas, cicCartuja, Universidad de Sevilla-Spanish National Scientific Council (CSIC), 41092 Seville, Spain.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: De la Rosa
authors.affiliation: Magnetic Resonance Center (CERM), Department of Chemistry, University of Florence, 50019 Sesto Fiorentino, Florence, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Turano
authors.affiliation: Instituto de Investigaciones Químicas, cicCartuja, Universidad de Sevilla-Spanish National Scientific Council (CSIC), 41092 Seville, Spain; idiazmoreno@us.es.
authors.forename: Irene
authors.initials: I
authors.lastname: Díaz-Moreno
mesh_terms: D045304:Cytochromes c / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008928:Mitochondria / Q000378:metabolism / Q000473:pathology*; D009154:Mutation; D018384:Oxidative Stress; D010544:Peroxidases / Q000378:metabolism; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011487:Protein Conformation; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction; D014443:Tyrosine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D017382:Reactive Oxygen Species; C075824:4-carboxymethylphenylalanine; D014443:Tyrosine; D010649:Phenylalanine; D045304:Cytochromes c; D010544:Peroxidases
keywords: cytochrome c; mitochondrial dysfunction; nuclear magnetic resonance; phosphorylation; respiratory supercomplexes