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PMID: 14614769 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Bax-induced cytochrome c release from mitochondria depends on alpha-helices-5 and -6.

The Biochemical journal ·Vol. 378 ·No. Pt 1 ·2004-02-15 ·Pages 247-55

Heimlich G, McKinnon AD, Bernardo K, Brdiczka D, Reed JC, Kain R, Krönke M, Jürgensmeier JM

Abstract

The pro-apoptotic protein Bax plays a key role in the mitochondrial signalling pathway. Upon induction of apoptosis, Bax undergoes a conformational change and translocates to mitochondrial membranes, where it inserts and mediates the release of cytochrome c from the intermembrane space into the cytosol. However, the domains of Bax that are essential for the induction of cytochrome c release are still elusive. Therefore various Bax deletion mutants were generated and expressed in Escherichia coli. The proteins were then purified in order to delineate the function of the transmembrane domain, the BH3 (Bcl-2 homology 3) domain and the putative pore-forming alpha-helices-5 and -6. These proteins were used to analyse the mechanism of Bax-induced cytochrome c release from mitochondria. None of the Bax proteins caused cytochrome c release merely through physical perturbation of the mitochondrial outer membrane. The alpha-helices-5 and -6 of Bax were shown to mediate the insertion of the protein into mitochondrial membranes and to be essential for the cytochrome c -releasing activity of Bax. In contrast, neither the transmembrane domain nor a functional BH3 domain is required for the Bax-mediated release of cytochrome c from mitochondria.

MeSH Terms
Animals Cytochromes c/metabolism Female Intracellular Membranes/metabolism Membrane Potentials Mice Mitochondria/metabolism,physiology,ultrastructure Mutation Protein Structure, Secondary Protein Structure, Tertiary Proto-Oncogene Proteins/chemistry,genetics,isolation & purification Proto-Oncogene Proteins c-bcl-2 Rats bcl-2-Associated X Protein
Chemicals
Bax protein, mouse Bax protein, rat Proto-Oncogene Proteins Proto-Oncogene Proteins c-bcl-2 bcl-2-Associated X Protein Cytochromes c
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Heimlich Gerd
Institute for Medical Microbiology, Immunology and Hygiene, University of Köln, 50935 Köln, Germany.
McKinnon Alastair D
Bernardo Katussevani
Brdiczka Dieter
Reed John C
Kain Renate
Krönke Martin
Jürgensmeier Juliane M
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
1470-8728
DOI
10.1042/BJ20031152
Published
2004-02-15
Pages
247-55
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1223939
Subset
IM
Grants
NIGMS NIH HHS · GM60554 · United States
External Links
PubMed source DOI: 10.1042/BJ20031152 PMC Full Text View journal details
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