Back to Search
Start Over
BAK/BAX activation and cytochrome c release assays using isolated mitochondria
- Source :
- Methods (San Diego, Calif.). 61(2)
- Publication Year :
- 2012
-
Abstract
- The mitochondrial pathway of apoptosis proceeds when the outer mitochondrial membrane (OMM) is compromised by the pro-apoptotic BCL-2 family members, BAK and BAX. Once activated, BAK and BAX form proteolipid pores in the OMM leading to mitochondrial outer membrane permeabilization (MOMP), and the release of inner membrane space proteins, such as cytochrome c, which promotes caspase activation. The use of isolated mitochondria has been instrumental to understanding the key interactions necessary to engage BAK and BAX activation, MOMP, and apoptosis. Furthermore, it is possible to biochemically define the relationships between BCL-2 family function and mitochondrial physiology using isolated systems. Our laboratory uses freshly isolated mitochondria from numerous sources to better understand BCL-2 family function and requirements for BAK and BAX activation. Here, we will discuss commonly used in vitro techniques to perform MOMP and cytochrome c release assays; and provide several key methodologies to implicate BAK and BAX activity in these processes.
- Subjects :
- Apoptosis
Mitochondria, Liver
Mitochondrion
Mitochondrial apoptosis-induced channel
General Biochemistry, Genetics and Molecular Biology
Article
Mice
Bcl-2-associated X protein
Inner membrane
Animals
Humans
Molecular Biology
bcl-2-Associated X Protein
biology
Cytochrome c
Bcl-2 family
Cytochromes c
Cell biology
Enzyme Activation
bcl-2 Homologous Antagonist-Killer Protein
Biochemistry
Gene Expression Regulation
Caspases
Mitochondrial Membranes
biology.protein
Biological Assay
Apoptosome
biological phenomena, cell phenomena, and immunity
Bcl-2 Homologous Antagonist-Killer Protein
Subjects
Details
- ISSN :
- 10959130
- Volume :
- 61
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Methods (San Diego, Calif.)
- Accession number :
- edsair.doi.dedup.....4567295dc0e2adbd2489f7792882de85