Induction of Apoptotic Program in Cell-Free Extracts: Requirement for dATP and Cytochrome c
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A cell-free system based on cytosols of normally growing cells is established that reproduces aspects of the apoptotic program in vitro. The apoptotic program is initiated by addition of dATP. Fractionation of cytosol yielded a 15 kDa protein that is required for in vitro apoptosis. The absorption spectrum and protein sequence revealed that this protein is cytochrome c. Elimination of cytochrome c from cytosol by immunodepletion, or inclusion of sucrose to stabilize mitochondria during cytosol preparation, diminished the apoptotic activity. Adding back cytochrome c to the cytochrome c-depleted extracts restored their apoptotic activity. Cells undergoing apoptosis in vivo showed increased release of cytochrome c to their cytosol, suggesting that mitochondria may function in apoptosis by releasing cytochrome c.
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MOMP directly promotes the cytosolic release of apoptogenic factors that normally reside in the mitochondrial intermembrane space, including cytochrome c (CYCS) and SMAC.
“MOMP directly promotes the cytosolic release of apoptogenic factors that normally reside in the mitochondrial intermembrane space [40, 44, 156]. These mitochondrial proteins include (but are not limited to) cytochrome c, somatic (CYCS), which usually operates as an electron shuttle in the mitochondrial respiratory chain [157–160], and diablo IAP-binding mitochondrial protein (DIABLO; also known as”
From Molecular mechanisms of cell death: recommendations of the nomenclature committee on cell death 2018. (2018) · cites this paper as [158]
If the pathological threshold is crossed, the electron transfer chain is disrupted, reactive oxygen species accumulate, the permeability transition pore opens, the outer mitochondrial membrane ruptures, cytochrome c is released, and an apoptotic program is activated.
“If the pathological threshold is crossed, the electron transfer chain is disrupted, reactive oxygen species accumulate, the permeability transition pore opens, the outer mitochondrial membrane ruptures, cytochrome c is released, and an apoptotic cell death program is activated [86,109].”
From Parvalbumin interneuron vulnerability and brain disorders (2021) · cites this paper as [86]
A substantial amount of cytochrome c and cytochrome c oxidase supports the high bioenergetic needs of PV-INs, but cytochrome c is also central to apoptosis induction.
“A substantial quantity of cytochrome c and cytochrome c oxidase supports the high bioenergetic needs of PV-INs [81,85], but comes at a price as cytochrome c is also central to the induction of apoptosis [86].”
From Parvalbumin interneuron vulnerability and brain disorders (2021) · cites this paper as [86]
MOMP and remodeling of mitochondrial membranes permit the release of apoptogenic factors such as cytochrome c and AIF from the intermembrane space into the cytosol.
“A key event in this process involves mitochondrial outer membrane permeabilization (MOMP) and the remodeling of mitochondrial membranes, permitting the release of apoptogenic factors such as cytochrome c (Liu et al., 1996) and apoptosis-inducing factor (AIF) (Susin et al., 1999) from the intermembrane space into the cytosol (Kroemer et al., 2007).”
From New insights into the role of mitochondria in aging: mitochondrial dynamics and more (2010) · cites this paper as Liu et al., 1996
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- Authors
- Liu, X., Kim, C. N., Yang, J., Jemmerson, R., & Wang, X.
- Affiliations
- Emory University (US)
- University of Minnesota (US)
- Corresponding author
- Wang, Xiaodong
- Venue
- Cell
- Publisher
- Elsevier BV
- Published
- 1996-07
- Volume
- 86
- Issue
- 1
- Pages
- 147-157
- ISSN
- 0092-8674 (ISSN-L)
- OpenAlex
- W2149967205
- Cited by
- 5,167
- References
- 49
- Citations by year
- 2026: 39 · 2025: 73 · 2024: 90 · 2023: 86 · 2022: 103 · 2021: 101 · 2020: 98 · 2019: 104 · 2018: 110 · 2017: 120 · 2016: 121 · 2015: 127 · 2014: 134 · 2013: 171 · 2012: 154
- FWCI
- 93.27
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Cell (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Keywords
- apoptosis, cytochrome c, dATP, cell-free extracts, cytosol, mitochondrial release, immunodepletion, mitochondrial stability
- MeSH terms
- Adenosine Triphosphate, Amino Acid Sequence, Antibodies, Monoclonal, Cell Extracts, Cell-Free System, Cysteine Endopeptidases, Cytochrome c Group, Cytosol, DNA, Enzyme Precursors, HeLa Cells, Humans, Molecular Sequence Data, Apoptosis, Caspases, Caspase 3
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