GO:0160007 glutathione import into mitochondrion: Mitochondrial Redox Homeostasis, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0160007 describes the biological process in which glutathione (GSH) is transported from the cytosol into the mitochondrial matrix.
• SLC25A39 is the essential mitochondrial carrier required for glutathione import in mammalian cells, and its loss causes mitochondrial GSH depletion, altered iron metabolism, and impaired heme synthesis [1,6].
• Mitochondrial glutathione import is autoregulated: SLC25A39 protein stability is controlled by mitochondrial GSH availability, creating a feedback loop that maintains mitochondrial redox homeostasis.
• Defective mitochondrial glutathione import promotes ferroptosis and has been linked to hepatic fibrotic injury, glioblastoma, and breast cancer metastasis [2,3,5].
• Mitochondrial GSH import supports integrated stress response signaling and enables breast cancer metastasis, making this pathway a potential therapeutic target.
• Researchers study GO:0160007 using CRISPR knockout, point-mutation, knock-in, overexpression models, and functional assays such as GSH quantification, ferroptosis sensitivity, and mitochondrial iron/heme measurements [1,4,6].
Description
Glutathione import into mitochondrion (GO:0160007) is the biological process that mediates the transport of glutathione from the cytosol into the mitochondrial matrix. Glutathione (GSH) is the most abundant non-enzymatic antioxidant in cells, and its mitochondrial pool is critical for protecting the organelle from oxidative damage, regulating iron-sulfur cluster biogenesis, and supporting heme synthesis [1,6]. The identification of SLC25A39 as the mitochondrial glutathione carrier established that this process is not merely passive diffusion but an active, carrier-mediated import step essential for mammalian cell viability. Mitochondrial glutathione import is now recognized as a central node in redox biology, iron metabolism, and cell death pathways [4,6]. Why does this matter for researchers? Mitochondrial GSH depletion caused by loss of SLC25A39 leads to impaired mitochondrial function, altered iron homeostasis, and increased sensitivity to ferroptosis, a form of iron-dependent lipid peroxidation-driven cell death [1,3,6]. The process is also autoregulated: SLC25A39 protein levels are controlled by mitochondrial GSH status, ensuring that import capacity matches the organelle's antioxidant demand. In cancer, mitochondrial glutathione import supports integrated stress response signaling and promotes breast cancer metastasis, highlighting its potential as a therapeutic target. In liver disease, the interaction between FUNDC1 and GPx4 governs hepatic ferroptosis and fibrotic injury through a mitophagy-dependent mechanism that intersects with mitochondrial GSH handling. This article provides a research-grade overview of GO:0160007, covering its definition, molecular machinery, key genes, regulatory mechanisms, disease relevance, and experimental methods including CRISPR-based models. All statements are grounded in the verified PubMed literature listed below.
glutathione import into mitochondrion At A Glance
| GO ID | GO:0160007 |
|---|---|
| GO term | glutathione import into mitochondrion |
| Ontology | biological_process |
| Synonym | none |
| Major function | Transport of glutathione from the cytosol into the mitochondrial matrix |
| Key carrier protein | SLC25A39 |
| Cellular location | Mitochondrial inner membrane / mitochondrial matrix |
| Regulatory feature | Autoregulatory control by mitochondrial GSH availability |
| Disease relevance | Ferroptosis, hepatic fibrotic injury, glioblastoma, breast cancer metastasis |
What Is GO:0160007?
GO:0160007 (glutathione import into mitochondrion) is defined as the process in which glutathione is transported from the cytosol into the mitochondrial matrix. This is a biological_process term in the Gene Ontology. The process requires the mitochondrial carrier protein SLC25A39 in mammalian cells, which facilitates the import of reduced glutathione (GSH) across the inner mitochondrial membrane. The term does not include glutathione synthesis, which occurs in the cytosol, nor does it include glutathione export or degradation; it specifically refers to the translocation step that delivers cytosolic GSH to the mitochondrial matrix [1,4].
Why Is glutathione import into mitochondrion Important in Cell Biology?
Mitochondrial glutathione import is essential for maintaining the mitochondrial redox environment, protecting against oxidative stress, and supporting iron-sulfur cluster and heme biosynthesis [1,6]. Loss of SLC25A39-mediated GSH import leads to mitochondrial GSH depletion, impaired mitochondrial function, and increased ferroptosis sensitivity, which has been implicated in liver disease, neurodegeneration, and cancer [1,3,5]. The autoregulatory feedback between mitochondrial GSH levels and SLC25A39 stability ensures that the import capacity adapts to the organelle's antioxidant needs, making this process a critical homeostatic mechanism. In cancer, mitochondrial GSH import supports integrated stress response signaling and enables metastasis, suggesting that targeting this pathway could have therapeutic benefit.
• Maintains mitochondrial redox homeostasis by supplying GSH to the mitochondrial matrix [1,7].
• Protects against ferroptosis, an iron-dependent form of cell death, by supporting GPx4-mediated lipid peroxide detoxification [3,5].
• Supports iron metabolism and heme synthesis through SLC25A39-dependent mitochondrial GSH import.
• Is autoregulated by mitochondrial GSH levels, ensuring adaptive control of import capacity.
• Promotes breast cancer metastasis via integrated stress response signaling.
• Is implicated in hepatic fibrotic injury through FUNDC1-GPx4 interactions and mitophagy.
• Represents a potential therapeutic target in glioblastoma, where SIRT3 targeting sensitizes cells to ferroptosis.
• Provides a mechanistic link between mitochondrial glutathione, coenzyme A metabolism, and thioredoxin reductase regulation.
• Enables researchers to study mitochondrial-specific antioxidant defense using CRISPR models [1,4].
• Connects mitochondrial GSH import to broader cellular stress responses and disease pathways [2,6].
What Happens During glutathione import into mitochondrion?
Recognition and binding of glutathione by SLC25A39
In simple terms: The carrier protein SLC25A39 on the mitochondrial inner membrane recognizes and binds glutathione from the cytosol.
SLC25A39 is a mitochondrial carrier protein localized to the inner mitochondrial membrane and is necessary for mitochondrial glutathione import in mammalian cells. It binds reduced glutathione (GSH) on the cytosolic side and undergoes conformational changes to translocate GSH into the mitochondrial matrix. Loss of SLC25A39 results in severe mitochondrial GSH depletion, demonstrating its essential role in this import process.
Translocation across the inner mitochondrial membrane
In simple terms: Glutathione is moved across the inner mitochondrial membrane into the matrix.
The import of glutathione into the mitochondrial matrix is mediated by SLC25A39, which functions as a carrier to transport GSH from the cytosol across the inner membrane. This translocation step is distinct from cytosolic glutathione synthesis and is required to maintain the mitochondrial GSH pool [1,4]. Mitochondrial GSH is critical for detoxifying reactive oxygen species and for the activity of glutathione-dependent enzymes such as GPx4 [3,7].
Autoregulatory feedback control of SLC25A39 stability
In simple terms: The amount of SLC25A39 protein is adjusted based on how much glutathione is already inside mitochondria.
Mitochondrial glutathione homeostasis is autoregulated: SLC25A39 protein stability is controlled by mitochondrial GSH availability, creating a feedback loop that matches import capacity to the organelle's antioxidant demand. When mitochondrial GSH levels are low, SLC25A39 is stabilized to increase import; when GSH levels are sufficient, SLC25A39 is degraded to prevent excess accumulation. This regulatory mechanism ensures that mitochondrial GSH import is dynamically adjusted to cellular needs.
Integration with iron metabolism and heme synthesis
In simple terms: Glutathione import affects how mitochondria handle iron and make heme.
SLC25A39 links mitochondrial GSH sensing with iron metabolism. Loss of SLC25A39-mediated GSH import leads to altered iron homeostasis and impaired heme synthesis, indicating that mitochondrial glutathione import is functionally connected to iron-sulfur cluster biogenesis and heme production. This integration is important for understanding how mitochondrial redox status influences broader metabolic pathways.
Role in ferroptosis and cell death regulation
In simple terms: Without glutathione import, mitochondria become vulnerable to ferroptosis, a type of cell death driven by lipid damage.
Mitochondrial glutathione import supports the activity of GPx4, which detoxifies lipid peroxides and prevents ferroptosis [3,5]. FUNDC1 interacts with GPx4 to govern hepatic ferroptosis and fibrotic injury through a mitophagy-dependent mechanism, linking mitochondrial GSH handling to cell death pathways. Targeting SIRT3 sensitizes glioblastoma to ferroptosis by promoting mitophagy and inhibiting SLC7A11, further highlighting the interplay between mitochondrial GSH import and ferroptosis regulation.
Key Genes Involved in GO:0160007 glutathione import into mitochondrion
The following genes and proteins are central to glutathione import into mitochondrion (GO:0160007) and its associated regulatory and disease pathways.
| Gene | Major Role | Research Relevance |
|---|---|---|
| SLC25A39 | Mitochondrial carrier essential for glutathione import into the mitochondrial matrix | Core transporter; knockout causes mitochondrial GSH depletion and ferroptosis sensitivity [1,6] |
| GPx4 | Glutathione peroxidase that detoxifies lipid peroxides using mitochondrial GSH | Interacts with FUNDC1 in hepatic ferroptosis and fibrotic injury |
| FUNDC1 | Mitophagy receptor that interacts with GPx4 to regulate ferroptosis | Links mitochondrial GSH handling to mitophagy-dependent cell death |
| SIRT3 | Mitochondrial deacetylase that regulates mitophagy and SLC7A11 | Targeting SIRT3 sensitizes glioblastoma to ferroptosis |
| SLC7A11 | Cystine/glutamate antiporter that supports glutathione synthesis | Inhibited by SIRT3 targeting in glioblastoma ferroptosis |
| Coenzyme A | Cofactor that protects against ferroptosis via CoAlation of thioredoxin reductase | Connects mitochondrial GSH import to thioredoxin reductase regulation |
| Thioredoxin reductase | Mitochondrial antioxidant enzyme regulated by CoAlation | CoAlation protects against ferroptosis |
| SLC25A39 (autoregulation) | Protein stability controlled by mitochondrial GSH availability | Feedback loop maintains mitochondrial GSH homeostasis |
| Integrated stress response (ISR) effectors | Signaling pathway enabled by mitochondrial GSH import | Promotes breast cancer metastasis |
| Heme synthesis enzymes | Iron metabolism and heme production linked to SLC25A39 | Loss of SLC25A39 impairs heme synthesis |
| Iron-sulfur cluster assembly proteins | Iron-sulfur cluster biogenesis connected to mitochondrial GSH | SLC25A39 links GSH sensing with iron metabolism |
| GPx4 (hepatic) | Glutathione peroxidase in liver | FUNDC1-GPx4 interaction governs hepatic ferroptosis |
| SIRT3 (glioblastoma) | Mitochondrial sirtuin | SIRT3 targeting promotes ferroptosis in glioblastoma |
| SLC7A11 (glioblastoma) | Cystine transporter | Inhibited by SIRT3 targeting, enhancing ferroptosis |
| CoA (mitochondrial) | Cofactor for CoAlation | Protects against ferroptosis via thioredoxin reductase CoAlation |
| Thioredoxin reductase (mitochondrial) | Antioxidant enzyme | CoAlation regulates its activity and ferroptosis protection |
How Is glutathione import into mitochondrion Regulated?
Mitochondrial glutathione import is regulated by an autoregulatory feedback mechanism in which SLC25A39 protein stability is controlled by mitochondrial GSH availability. When mitochondrial GSH levels decline, SLC25A39 is stabilized to increase glutathione import; when GSH levels are sufficient, SLC25A39 is degraded to prevent excess accumulation. This feedback loop ensures that mitochondrial GSH import capacity is matched to the organelle's antioxidant demand. Additionally, mitochondrial GSH import supports integrated stress response (ISR) signaling, which can influence cancer cell metastasis. The process is also linked to iron metabolism, as SLC25A39 connects mitochondrial GSH sensing with iron homeostasis and heme synthesis.
glutathione import into mitochondrion and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| SLC25A39 | Breast cancer metastasis via ISR signaling | SLC25A39 knockout breast cancer cell lines; metastasis assays |
| FUNDC1 | Hepatic ferroptosis and fibrotic injury | FUNDC1 knockout hepatocytes; liver fibrosis models |
| GPx4 | Hepatic ferroptosis and fibrotic injury | GPx4 knockout or point-mutation models; ferroptosis assays |
| SIRT3 | Glioblastoma ferroptosis sensitivity | SIRT3 knockout glioblastoma cells; ferroptosis induction |
| SLC7A11 | Glioblastoma ferroptosis sensitivity | SLC7A11 knockout or overexpression glioblastoma models |
Mitochondrial glutathione import in cancer metastasis
Mitochondrial glutathione import enables breast cancer metastasis via integrated stress response signaling. SLC25A39-mediated GSH import supports the ISR pathway, which promotes metastatic dissemination. This suggests that targeting mitochondrial GSH import could be a therapeutic strategy to limit cancer spread.
Ferroptosis and hepatic fibrotic injury
FUNDC1 interacts with GPx4 to govern hepatic ferroptosis and fibrotic injury through a mitophagy-dependent manner. Mitochondrial glutathione import is essential for GPx4 activity, which detoxifies lipid peroxides and prevents ferroptosis. Disruption of this process leads to hepatic ferroptosis and fibrotic injury, highlighting the importance of mitochondrial GSH import in liver disease.
Glioblastoma and ferroptosis sensitivity
Targeting SIRT3 sensitizes glioblastoma to ferroptosis by promoting mitophagy and inhibiting SLC7A11. This sensitization is linked to mitochondrial glutathione import, as SLC7A11 supports glutathione synthesis and mitochondrial GSH import maintains the mitochondrial antioxidant pool. The interplay between SIRT3, SLC7A11, and mitochondrial GSH import suggests new therapeutic approaches for glioblastoma.
Iron metabolism and heme synthesis disorders
SLC25A39 links mitochondrial GSH sensing with iron metabolism. Loss of SLC25A39-mediated glutathione import leads to altered iron homeostasis and impaired heme synthesis, which may contribute to disorders of iron metabolism and mitochondrial function. This connection underscores the broader metabolic importance of mitochondrial glutathione import.
From glutathione import into mitochondrion-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of SLC25A39 impair mitochondrial glutathione import? | SLC25A39 knockout cell lines (e.g., HEK293T, HeLa) |
| How does mitochondrial GSH availability regulate SLC25A39 stability? | SLC25A39 tagged knock-in with degron or reporter; GSH modulation |
| Does SLC25A39-mediated GSH import affect iron metabolism and heme synthesis? | SLC25A39 knockout cells; iron and heme quantification |
| Can mitochondrial GSH import promote breast cancer metastasis? | SLC25A39 knockout or overexpression in breast cancer cells; metastasis models |
| How does FUNDC1-GPx4 interaction regulate hepatic ferroptosis? | FUNDC1 knockout or point-mutation hepatocytes; ferroptosis assays |
| Does SIRT3 targeting sensitize glioblastoma to ferroptosis via SLC7A11? | SIRT3 knockout or overexpression glioblastoma cells; ferroptosis assays |
How to Study the glutathione import into mitochondrion Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Mitochondrial GSH quantification | Glutathione levels in mitochondrial matrix | Assessing SLC25A39 knockout effects |
| Genetically encoded GSH sensors | Real-time mitochondrial GSH dynamics | Monitoring autoregulation of SLC25A39 |
| Ferroptosis assays | Lipid peroxidation and cell viability | Testing sensitivity to ferroptosis inducers [3,5] |
| Iron quantification (ICP-MS) | Total and mitochondrial iron levels | Evaluating SLC25A39 link to iron metabolism |
| Heme quantification | Heme synthesis capacity | Assessing mitochondrial function in knockout cells |
| CRISPR knockout | Loss-of-function phenotypes | Determining essentiality of SLC25A39 |
| CRISPR knock-in (tagged) | Protein localization and stability | Studying SLC25A39 autoregulation |
| Overexpression | Gain-of-function effects | Testing sufficiency of SLC25A39 in GSH import |
Quantification of mitochondrial glutathione
Mitochondrial glutathione levels can be measured using biochemical assays or genetically encoded fluorescent sensors to assess the impact of SLC25A39 loss or modulation [1,4]. These methods are essential for confirming that GO:0160007 is disrupted in knockout or point-mutation models.
Ferroptosis sensitivity assays
Ferroptosis sensitivity is assessed by treating cells with ferroptosis inducers (e.g., erastin, RSL3) and measuring lipid peroxidation, cell viability, and GPx4 activity [3,5]. These assays link mitochondrial glutathione import to cell death pathways [3,5].
Iron and heme quantification
Iron and heme levels are measured using colorimetric assays, inductively coupled plasma mass spectrometry (ICP-MS), or fluorescent probes to evaluate the connection between SLC25A39 and iron metabolism. These methods help determine how mitochondrial GSH import affects heme synthesis.
CRISPR-based genetic models
CRISPR knockout, point-mutation, knock-in, and overexpression models are used to dissect the function of SLC25A39 and related genes in mitochondrial glutathione import [1,4,6]. These models enable causal testing of gene function in disease-relevant contexts [2,3,5].
How CRISPR Can Be Used to Study GO:0160007 glutathione import into mitochondrion
Knockout
CRISPR knockout of SLC25A39 in mammalian cells abolishes mitochondrial glutathione import, leading to mitochondrial GSH depletion, impaired iron metabolism, and increased ferroptosis sensitivity [1,6]. Knockout models are essential for establishing the causal role of SLC25A39 in GO:0160007.
Point Mutation
Point mutations in SLC25A39 can be introduced to dissect the residues required for glutathione binding and transport, as well as to model disease-associated variants [1,4]. Such models help distinguish between loss-of-function and gain-of-function mechanisms.
Knock-in
Knock-in of tagged SLC25A39 (e.g., HA, FLAG, or degron tags) allows for precise monitoring of protein stability, localization, and autoregulatory degradation in response to mitochondrial GSH levels. This approach is valuable for studying the feedback loop controlling mitochondrial glutathione import.
Overexpression
Overexpression of SLC25A39 increases mitochondrial glutathione import capacity and can protect cells from ferroptosis, while overexpression in cancer models may promote metastasis via ISR signaling [1,2]. Overexpression models are useful for testing sufficiency and therapeutic potential.
How EDITGENE Supports glutathione import into mitochondrion Research
Researchers studying glutathione import into mitochondrion-related genes often need to determine whether a candidate gene is causally involved in mitochondrial GSH homeostasis, ferroptosis, or disease progression. EDITGENE provides comprehensive CRISPR-based services to generate knockout, point-mutation, knock-in, and overexpression cell models, as well as CRISPR library screening and bioinformatics support, enabling rigorous functional studies of GO:0160007 and its associated pathways.
Contact EDITGENE today to design your custom CRISPR model for glutathione import into mitochondrion research.
Frequently Asked Questions About glutathione import into mitochondrion
What is GO:0160007?
GO:0160007 is the Gene Ontology term for glutathione import into mitochondrion, the process in which glutathione is transported from the cytosol into the mitochondrial matrix.
What genes are involved in glutathione import into mitochondrion?
SLC25A39 is the essential mitochondrial carrier for glutathione import in mammalian cells. Other genes such as GPx4, FUNDC1, SIRT3, and SLC7A11 are functionally linked to mitochondrial glutathione metabolism and ferroptosis [3,5].
Why is mitochondrial glutathione import important?
It maintains mitochondrial redox homeostasis, protects against oxidative stress and ferroptosis, supports iron metabolism and heme synthesis, and enables integrated stress response signaling in cancer [1,2,6].
How is SLC25A39 regulated?
SLC25A39 protein stability is autoregulated by mitochondrial glutathione availability, creating a feedback loop that matches import capacity to antioxidant demand.
What diseases are associated with defective mitochondrial glutathione import?
Defective import is linked to hepatic ferroptosis and fibrotic injury, glioblastoma ferroptosis sensitivity, breast cancer metastasis, and disorders of iron metabolism [2,3,5,6].
How can I study glutathione import into mitochondrion in the lab?
You can use CRISPR knockout, point-mutation, knock-in, and overexpression models combined with mitochondrial GSH quantification, ferroptosis assays, and iron/heme measurements [1,4,6].
What is the role of SLC25A39 in ferroptosis?
SLC25A39-mediated glutathione import supports GPx4 activity, which detoxifies lipid peroxides and prevents ferroptosis [1,3].
Does mitochondrial glutathione import affect cancer metastasis?
Yes, mitochondrial glutathione import enables breast cancer metastasis via integrated stress response signaling.
How does FUNDC1 relate to mitochondrial glutathione import?
FUNDC1 interacts with GPx4 to govern hepatic ferroptosis and fibrotic injury through a mitophagy-dependent mechanism, linking mitochondrial GSH handling to cell death.
Can CRISPR be used to study GO:0160007?
Yes, CRISPR knockout, point-mutation, knock-in, and overexpression models are widely used to dissect the function of SLC25A39 and related genes in mitochondrial glutathione import [1,4,6].
Conclusion
Glutathione import into mitochondrion (GO:0160007) is a fundamental biological process required for mitochondrial redox homeostasis, iron metabolism, and protection against ferroptosis [1,6]. The identification of SLC25A39 as the essential carrier and the discovery of its autoregulatory control have provided mechanistic insights into how mitochondria maintain their glutathione pool [1,4]. Dysregulation of this process is implicated in cancer metastasis, hepatic fibrotic injury, and glioblastoma, making it a promising therapeutic target [2,3,5]. Researchers can leverage CRISPR-based models, including knockout, point-mutation, knock-in, and overexpression cell lines, to dissect the molecular details of GO:0160007 and its disease relevance. EDITGENE offers comprehensive services to support these studies, from model generation to library screening and bioinformatics analysis.
References
- 1. Wang Y et al.. 2021. SLC25A39 is necessary for mitochondrial glutathione import in mammalian cells.. Nature 599(7883):136-140 PMID: 34707288
- 2. Yeh HW et al.. 2025. Mitochondrial Glutathione Import Enables Breast Cancer Metastasis via Integrated Stress Response Signaling.. Cancer Discov 15(12):2437-2449 PMID: 40736010
- 3. Bi Y et al.. 2024. FUNDC1 interacts with GPx4 to govern hepatic ferroptosis and fibrotic injury through a mitophagy-dependent manner.. J Adv Res 55:45-60 PMID: 36828120
- 4. Liu Y et al.. 2023. Autoregulatory control of mitochondrial glutathione homeostasis.. Science 382(6672):820-828 PMID: 37917749
- 5. Li X et al.. 2024. Targeting SIRT3 sensitizes glioblastoma to ferroptosis by promoting mitophagy and inhibiting SLC7A11.. Cell Death Dis 15(2):168 PMID: 38395990
- 6. Chen X et al.. 2024. SLC25A39 links mitochondrial GSH sensing with iron metabolism.. Mol Cell 84(4):616-618 PMID: 38364779
- 7. Ribas V et al.. 2014. Glutathione and mitochondria.. Front Pharmacol 5:151 PMID: 25024695
- 8. Lin CC et al.. 2025. Coenzyme A protects against ferroptosis via CoAlation of mitochondrial thioredoxin reductase.. J Clin Invest 135(19) PMID: 40694424
