Bax releases cytochrome c preferentially from a complex between porin and adenine nucleotide translocator. Hexokinase activity suppresses this effect.
The mechanism by which external Bax releases cytochrome c is still controversial and may also depend on the type of mitochondria and the actual localisation of cytochrome c. Outer membrane porin acquires high binding affinity for hexokinase by interacting with the adenine nucleotide translocator (ANT) in the contact sites. (I) The hexokinase protein was thus used as a tool to isolate the contact site forming complex between outer membrane porin and inner membrane ANT from a TritonX100 extract of brain membranes. (II) A significant amount of cytochrome c was co-purified with the isolated hexokinase porin ANT complexes that were reconstituted in phospholipid vesicles. Bax-AC released the endogenous cytochrome c from the vesicles without forming unspecific pores. This was shown by loading the vesicles with malate that was not liberated by Bax-AC. (III) The Bax-AC effect was dependent on a specific association of cytochrome c with the porin ANT complex, as dissociation of the complex by bongkrekate abolished the Bax dependent cytochrome c liberation. (IV) The Bax-AC effect was as well suppressed by hexokinase phosphorylating glucose.
Vyssokikh
Miklail Y
MY
A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University.
Zorova
Ljubava
L
Zorov
Dmitry
D
Heimlich
Gerd
G
Jürgensmeier
Juliane J
JJ
Brdiczka
Dieter
D
D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D003574:Cytochrome c Group / Q000378:metabolism*; D006593:Hexokinase / Q000378:metabolism*; D046911:Macromolecular Substances; D008928:Mitochondria / Q000378:metabolism; D000226:Mitochondrial ADP, ATP Translocases / Q000378:metabolism*; D018272:Porins / Q000378:metabolism*; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2; D050994:Voltage-Dependent Anion Channels; D051028:bcl-2-Associated X Protein
D003574:Cytochrome c Group; D046911:Macromolecular Substances; D018272:Porins; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D050994:Voltage-Dependent Anion Channels; D051028:bcl-2-Associated X Protein; D000226:Mitochondrial ADP, ATP Translocases; D006593:Hexokinase