title: Analysis of pro-apoptotic protein trafficking to and from mitochondria.
abstract: Mitochondria play a key role in cell death and its regulation. The permeabilization of the outer mitochondrial membrane which is mainly controlled by proteins of the BCL-2 family, is a key event that can be directly induced by p53 and results in the release of pro-apoptotic factors to the cytosol, such as cytochrome c, second mitochondria derived activator of caspases/direct inhibitor-of-apoptosis (IAP) binding protein with low pI (SMAC/Diablo), Omi serine protease (Omi/HtrA2), apoptosis inducing factor (AIF), or endonuclease G (Endo-G). Hence, the determination of subcellular localization of these proteins is extremely important to predict cell fate and elucidate the specific mechanism of apoptosis. Here we describe the procedures that can be used to study the subcellular location of different pro-apoptotic proteins to be used in basic cell biology and toxicology studies.
authors.affiliation: CNC - Center for Neuroscience and Cell Biology, Largo Marquês de Pombal, University of Coimbra, 3004-517, Coimbra, Portugal.
authors.forename: Ignacio
authors.initials: I
authors.lastname: Vega-Naredo
authors.forename: Teresa
authors.initials: T
authors.lastname: Cunha-Oliveira
authors.forename: Teresa L
authors.initials: TL
authors.lastname: Serafim
authors.forename: Vilma A
authors.initials: VA
authors.lastname: Sardao
authors.forename: Paulo J
authors.initials: PJ
authors.lastname: Oliveira
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D002458:Cell Fractionation; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods*; D047468:Immunoprecipitation; D051379:Mice; D008928:Mitochondria / Q000378:metabolism*; D021381:Protein Transport; D051381:Rats; D016707:Tissue Fixation
chemical_list: D051017:Apoptosis Regulatory Proteins; D005456:Fluorescent Dyes