GO:0005347 ATP transmembrane transporter activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0005347 ATP transmembrane transporter activity describes the molecular function that enables the transfer of ATP across a membrane.
• This activity is essential for mitochondrial energy export, ATP-dependent ion pumps, ABC transporter signaling, and extracellular purinergic signaling [1,2,4].
• Key proteins include mitochondrial ATP/ADP carriers (SLC25A4, SLC25A5, SLC25A6), the Na,K-ATPase (ATP1A1), and ABC transporters such as CFTR and P-glycoprotein (ABCB1) [1,2,4,7].
• Dysregulation of ATP transport contributes to mitochondrial permeability transition, cystic fibrosis, neurodegeneration, and drug resistance in cancer [1,2,7].
• CRISPR knockout, point mutation, knock-in, and overexpression models are powerful tools to dissect the causal roles of ATP transporter genes [3,6].
• EDITGENE provides end-to-end CRISPR services, including library screening and bioinformatics, to accelerate research on ATP transmembrane transporter activity.
Description
ATP transmembrane transporter activity (GO:0005347) is a molecular function that enables the movement of adenosine triphosphate (ATP) across biological membranes. This activity is fundamental to cellular energy distribution, as ATP generated in mitochondria must be exported to the cytosol to fuel numerous energy-consuming processes. Additionally, ATP transport across the plasma membrane is critical for extracellular purinergic signaling, which regulates neurotransmission, inflammation, and immune responses. The dysfunction of ATP transporters is linked to a wide range of human diseases, including mitochondrial myopathies, cystic fibrosis, and cancer drug resistance [1,2,7]. Understanding the molecular mechanisms and regulation of ATP transmembrane transporter activity is therefore of paramount importance for both basic biology and therapeutic development. Researchers studying this function employ a variety of experimental approaches, from biochemical assays to CRISPR-based genetic models, to uncover how specific genes contribute to ATP transport and how their dysregulation leads to disease [3,6].
ATP transmembrane transporter activity At A Glance
| GO ID | GO:0005347 |
|---|---|
| GO term | ATP transmembrane transporter activity |
| Ontology | molecular_function |
| Synonym | None |
| Major function | Transfer of ATP across a membrane |
| Definition | Enables the transfer of ATP, adenosine triphosphate, from one side of a membrane to the other. |
| Related transporters | Mitochondrial ATP/ADP carriers (SLC25A4/5/6), Na,K-ATPase (ATP1A1), ABC transporters (CFTR, ABCB1) |
| Associated diseases | Mitochondrial myopathies, cystic fibrosis, cancer drug resistance, neurodegeneration |
What Is GO:0005347?
According to the Gene Ontology, GO:0005347 ATP transmembrane transporter activity is defined as the function that enables the transfer of ATP, adenosine triphosphate, from one side of a membrane to the other. This activity is distinct from ATP synthesis or hydrolysis; it specifically describes the translocation of intact ATP molecules across lipid bilayers. It is a molecular function that can be carried out by various proteins, including mitochondrial carriers, ABC transporters, and ion pumps, each utilizing distinct mechanisms such as facilitated diffusion, primary active transport, or co-transport [1,2,4].
Why Is ATP transmembrane transporter activity Important in Cell Biology?
ATP transmembrane transporter activity is central to cellular energy homeostasis and intercellular communication. It ensures that ATP produced in mitochondria is available to the cytosol and that extracellular ATP can act as a signaling molecule. Defects in ATP transport are implicated in a spectrum of disorders, from rare mitochondrial diseases to common pathologies like cystic fibrosis and cancer [1,2,7]. Moreover, many drugs and natural products interact with ATP transporters, affecting their pharmacokinetics and efficacy. Thus, studying this activity is crucial for understanding basic physiology and for developing targeted therapies.
• Maintains cellular energy balance by exporting mitochondrial ATP to the cytosol.
• Enables extracellular ATP signaling, which regulates neurotransmission, inflammation, and immunity.
• Supports the function of ATP-dependent ion pumps, such as the Na,K-ATPase, which maintains membrane potential.
• Modulates the activity of ABC transporters, including CFTR and P-glycoprotein, which are involved in cystic fibrosis and multidrug resistance [2,7].
• Dysregulation leads to mitochondrial permeability transition, a key event in cell death and ischemia-reperfusion injury.
• ATP transporters influence drug absorption, distribution, and excretion, impacting therapeutic outcomes.
• Mutations in ATP transporter genes cause diseases such as mitochondrial DNA depletion syndromes and cystic fibrosis [1,2].
• ATP transport is essential for proper brain function, and its impairment is linked to neurodegenerative diseases like ALS.
• Targeting ATP transporters can overcome drug resistance in cancer and improve treatment efficacy.
• CRISPR-based models of ATP transporter genes facilitate the discovery of novel therapeutic targets [3,6].
What Happens During ATP transmembrane transporter activity?
Substrate Recognition and Binding
In simple terms: The transporter first grabs ATP on one side of the membrane.
ATP transmembrane transporters specifically recognize and bind ATP. For example, mitochondrial ATP/ADP carriers (SLC25A4, SLC25A5, SLC25A6) bind ATP from the mitochondrial matrix and exchange it for ADP from the intermembrane space. The binding site involves conserved positively charged residues that interact with the phosphate groups of ATP. In ABC transporters like CFTR, ATP binds to nucleotide-binding domains (NBDs), inducing conformational changes [2,5]. The specificity for ATP over other nucleotides is determined by the shape and charge of the binding pocket.
Conformational Change and Translocation
In simple terms: The transporter changes shape to move ATP across the membrane.
Upon ATP binding, transporters undergo conformational changes that allow ATP to pass through the membrane. Mitochondrial carriers alternate between two conformations: one open to the matrix and one open to the intermembrane space, facilitating ATP/ADP exchange. ABC transporters use the energy of ATP binding and hydrolysis to drive conformational cycles that transport substrates, although some, like CFTR, function as channels [2,5]. The Na,K-ATPase, a P-type ATPase, undergoes phosphorylation and dephosphorylation cycles to transport ions, but it also binds ATP for its catalytic cycle.
Energy Coupling and Regulation
In simple terms: Some transporters use energy to pump ATP, while others just let it flow.
ATP transport can be passive or active. Mitochondrial ATP/ADP exchange is driven by the electrochemical gradient across the inner membrane. In contrast, ABC transporters like P-glycoprotein (ABCB1) hydrolyze ATP to actively efflux drugs and other substrates, but they also transport ATP itself in some contexts. The Na,K-ATPase hydrolyzes ATP to pump ions against their gradients, and ATP binding is essential for its function. Regulation occurs via post-translational modifications, such as phosphorylation, and through interactions with regulatory proteins.
Release and Recycling
In simple terms: After moving ATP, the transporter resets to start again.
Following translocation, ATP is released on the other side of the membrane. The transporter then returns to its initial conformation to complete the cycle. For mitochondrial carriers, the exchange of ATP for ADP is tightly coupled to avoid futile cycling. ABC transporters hydrolyze ATP to reset their conformation, ensuring processive transport [2,5]. The Na,K-ATPase dephosphorylates to return to its E1 state, ready for another round of ion transport. This recycling is critical for maintaining continuous ATP transport.
Key Genes Involved in GO:0005347 ATP transmembrane transporter activity
The following genes encode proteins that exhibit ATP transmembrane transporter activity or are directly involved in ATP transport across membranes.
| Gene | Major Role | Research Relevance |
|---|---|---|
| SLC25A4 | Mitochondrial ATP/ADP carrier (ANT1) | Mutations cause mitochondrial DNA depletion and myopathy |
| SLC25A5 | Mitochondrial ATP/ADP carrier (ANT2) | Highly expressed in proliferating cells; target in cancer |
| SLC25A6 | Mitochondrial ATP/ADP carrier (ANT3) | X-linked; involved in energy metabolism |
| ATP1A1 | Na,K-ATPase alpha-1 subunit | Maintains ion gradients; target for cardiotonic steroids |
| ATP1A2 | Na,K-ATPase alpha-2 subunit | Mutations cause familial hemiplegic migraine |
| ATP1A3 | Na,K-ATPase alpha-3 subunit | Mutations cause rapid-onset dystonia-parkinsonism |
| ATP1B1 | Na,K-ATPase beta-1 subunit | Regulates pump assembly and function |
| CFTR | ABC transporter; chloride channel | Mutations cause cystic fibrosis; ATP binding regulates gating [2,5] |
| ABCB1 | P-glycoprotein; multidrug transporter | ATP-dependent efflux; involved in drug resistance |
| ABCC1 | Multidrug resistance protein 1 | ATP-dependent transport of drugs and organic anions |
| ABCG2 | Breast cancer resistance protein | ATP-dependent efflux; affects drug bioavailability |
| VDAC1 | Voltage-dependent anion channel | Allows ATP/ADP flux across outer mitochondrial membrane |
| VDAC2 | Voltage-dependent anion channel 2 | Regulates mitochondrial ATP release |
| VDAC3 | Voltage-dependent anion channel 3 | Less characterized; may modulate ATP transport |
| SLC25A31 | Mitochondrial ATP/ADP carrier (ANT4) | Testis-specific; role in sperm function |
| SLC25A17 | Peroxisomal ATP transporter | Transports ATP into peroxisomes |
| SLC25A42 | Mitochondrial CoA/ATP carrier | Linked to metabolic disorders |
How Is ATP transmembrane transporter activity Regulated?
ATP transmembrane transporter activity is regulated at multiple levels. Transcriptional regulation controls the expression of transporter genes in response to energy demand and stress. Post-translational modifications, such as phosphorylation by kinases like PKA and PKC, modulate transporter activity [2,4]. For example, CFTR channel gating is regulated by phosphorylation of its regulatory domain and ATP binding to its nucleotide-binding domains [2,5]. The Na,K-ATPase is regulated by hormones (e.g., insulin) and intracellular calcium. Additionally, the mitochondrial permeability transition pore (mPTP) regulates ATP transport across the inner mitochondrial membrane and is modulated by cyclophilin D. In cancer cells, P-glycoprotein (ABCB1) activity is regulated by signaling pathways such as PI3K/Akt, contributing to drug resistance.
ATP transmembrane transporter activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| SLC25A4 | Mitochondrial DNA depletion syndrome 12 (cardiomyopathic type) | Knockout mouse, patient-derived iPSCs |
| CFTR | Cystic fibrosis | CFTR knockout pig, human bronchial epithelial cells |
| ABCB1 | Multidrug resistance in cancer | ABCB1 knockout cancer cell lines, xenografts |
| ATP1A2 | Familial hemiplegic migraine | Knock-in mouse with patient mutation |
| ATP1A3 | Rapid-onset dystonia-parkinsonism | Knock-in mouse, patient iPSC-derived neurons |
Mitochondrial Diseases and Permeability Transition
Mutations in mitochondrial ATP/ADP carriers (SLC25A4, SLC25A5, SLC25A6) cause mitochondrial DNA depletion syndromes, which manifest as myopathy, cardiomyopathy, and hepatopathy. The mitochondrial permeability transition pore (mPTP) is a key mediator of cell death; its opening leads to ATP depletion and necrosis, contributing to ischemia-reperfusion injury and neurodegeneration. Targeting mPTP components, including ATP transporters, is a therapeutic strategy for these conditions.
Cystic Fibrosis and ABC Transporter Dysfunction
Cystic fibrosis is caused by mutations in CFTR, an ABC transporter that functions as a chloride channel. ATP binding and hydrolysis are essential for CFTR gating, and mutations that impair ATP interaction lead to channel dysfunction [2,5]. CFTR also transports ATP, which affects extracellular purinergic signaling in the airways, contributing to disease pathology. Modulators that correct CFTR folding and gating have revolutionized treatment, but further understanding of ATP-dependent mechanisms is needed.
Cancer Drug Resistance and Neurodegeneration
Overexpression of ABCB1 (P-glycoprotein) and other ABC transporters confers multidrug resistance in cancer by actively effluxing chemotherapeutic agents in an ATP-dependent manner. In amyotrophic lateral sclerosis (ALS), increased ATP production and P-glycoprotein activity at the blood-brain barrier alter drug transport, potentially affecting treatment response. Modulating ATP transporter activity could enhance drug delivery to the brain and improve outcomes in neurodegenerative diseases.
From ATP transmembrane transporter activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of SLC25A4 impair mitochondrial ATP export? | SLC25A4 knockout HeLa or HEK293 cells |
| Does a specific CFTR mutation affect ATP binding? | CFTR point-mutation knock-in (e.g., G551D) in CFBE41o- cells |
| Can overexpression of ABCB1 confer drug resistance? | ABCB1 overexpression in cancer cell lines (e.g., MCF-7) |
| What is the role of ATP1A1 in ion homeostasis? | ATP1A1 knockout or knockdown in renal epithelial cells |
| How does VDAC1 regulate ATP flux? | VDAC1 knockout mouse embryonic fibroblasts |
| Can CRISPR screening identify novel ATP transporters? | Genome-wide CRISPR knockout library in ATP-dependent reporter cells |
How to Study the ATP transmembrane transporter activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Radioactive ATP transport assay | ATP flux across membranes | Mitochondrial carrier activity |
| ATPase activity assay | ATP hydrolysis rate | ABC transporter function [2,7] |
| Patch-clamp electrophysiology | Ion channel currents and ATP sensitivity | CFTR gating [2,5] |
| Live-cell ATP imaging | Intracellular/extracellular ATP dynamics | Mitochondrial ATP export |
| CRISPR knockout screen | Gene essentiality for ATP transport | Discovery of novel transporters |
| RNA-seq | Transcriptional changes in transporter genes | Response to metabolic stress |
| Cryo-EM | High-resolution structure of transporters | ATP binding site mapping |
| Molecular dynamics simulation | ATP translocation pathways | Mutation impact prediction |
Biochemical Transport Assays
ATP transport can be measured using radioactive ATP (e.g., [α-32P]ATP) or fluorescent ATP analogs in isolated membrane vesicles or reconstituted proteoliposomes [1,4]. For mitochondrial carriers, exchange activity is assayed by monitoring ATP/ADP exchange using stopped-flow kinetics. For ABC transporters, ATPase activity is measured by colorimetric or luminescent assays [2,7]. These methods provide direct kinetic parameters and substrate specificity.
Electrophysiology and Imaging
Patch-clamp electrophysiology is used to study ATP-dependent channels like CFTR, measuring single-channel currents and ATP sensitivity [2,5]. Live-cell imaging with fluorescent ATP sensors (e.g., ATeam, PercevalHR) allows real-time monitoring of ATP transport across membranes. Mitochondrial ATP export can be visualized using targeted luciferase or fluorescent probes. These techniques reveal dynamic changes in intracellular and extracellular ATP levels.
Genetic and CRISPR Screens
CRISPR knockout, knock-in, and overexpression models enable functional dissection of ATP transporter genes [3,6]. Genome-wide CRISPR screens with ATP-dependent reporters can identify novel transporters and regulators. For example, a screen for genes affecting extracellular ATP levels can uncover new players in purinergic signaling. These approaches are complemented by RNA-seq and proteomics to assess expression changes.
Structural and Computational Approaches
Cryo-EM and X-ray crystallography provide high-resolution structures of ATP transporters, revealing ATP binding sites and conformational states. Molecular dynamics simulations can model ATP translocation and predict mutation effects. These methods guide the design of point mutations to test mechanistic hypotheses. Combined with biochemical assays, they offer a comprehensive understanding of ATP transport.
How CRISPR Can Be Used to Study GO:0005347 ATP transmembrane transporter activity
Knockout
CRISPR knockout of ATP transporter genes (e.g., SLC25A4, CFTR, ABCB1) creates cell models to study loss-of-function phenotypes. For example, SLC25A4 knockout cells exhibit impaired mitochondrial ATP export and reduced cytosolic ATP levels. CFTR knockout cells are used to study chloride transport and ATP-dependent gating. ABCB1 knockout cancer cells become sensitive to chemotherapeutic drugs, confirming its role in multidrug resistance. These models are essential for validating gene function and identifying compensatory mechanisms.
Point Mutation
CRISPR point mutation (base editing or homology-directed repair) introduces specific disease-associated mutations into ATP transporter genes. For instance, the CFTR G551D mutation, which impairs ATP-dependent gating, can be knocked into airway epithelial cells to study cystic fibrosis [2,5]. Similarly, mutations in ATP1A2 (e.g., W887R) linked to hemiplegic migraine can be modeled in neurons. These models provide insights into how single amino acid changes affect ATP binding, transport kinetics, and disease pathology.
Knock-in
Knock-in of tagged ATP transporters (e.g., GFP or HA tags) allows real-time visualization and purification of transporter complexes. For example, knocking in a fluorescent tag on SLC25A4 enables live-cell imaging of mitochondrial ATP carrier dynamics. Tagged CFTR can be used to study its trafficking and interaction with ATP. Knock-in of reporter genes (e.g., luciferase) under the control of transporter promoters allows monitoring of expression changes in response to metabolic cues.
Overexpression
CRISPR activation (CRISPRa) or lentiviral overexpression of ATP transporter genes is used to study gain-of-function effects. Overexpression of ABCB1 in cancer cells confers drug resistance, providing a model to test P-glycoprotein inhibitors. Overexpression of SLC25A5 (ANT2) enhances mitochondrial ATP export and supports proliferation. These models are valuable for screening compounds that modulate ATP transport and for understanding how increased transporter activity contributes to disease.
How EDITGENE Supports ATP transmembrane transporter activity Research
Researchers studying ATP transmembrane transporter activity-related genes often need to determine whether a candidate gene is causally involved in ATP transport, how specific mutations affect transporter function, and whether targeting these genes can reverse disease phenotypes. EDITGENE provides a comprehensive suite of CRISPR services to address these questions with precision and efficiency.
Contact EDITGENE today to design your custom CRISPR model for ATP transmembrane transporter activity research.
Frequently Asked Questions About ATP transmembrane transporter activity
What is ATP transmembrane transporter activity?
ATP transmembrane transporter activity (GO:0005347) is a molecular function that enables the transfer of ATP across a membrane, as defined by the Gene Ontology.
What genes are involved in ATP transmembrane transporter activity?
Key genes include SLC25A4, SLC25A5, SLC25A6 (mitochondrial ATP/ADP carriers), ATP1A1 (Na,K-ATPase), CFTR, and ABCB1 (ABC transporters) [1,2,4,7].
How is ATP transported across the mitochondrial membrane?
Mitochondrial ATP/ADP carriers (SLC25A4/5/6) exchange ATP from the matrix for ADP from the intermembrane space, driven by the electrochemical gradient.
What diseases are associated with defects in ATP transport?
Defects are linked to mitochondrial DNA depletion syndromes, cystic fibrosis, cancer drug resistance, and neurodegenerative diseases like ALS [1,2,7].
How can I study ATP transmembrane transporter activity in the lab?
Common methods include radioactive ATP transport assays, patch-clamp electrophysiology, live-cell ATP imaging, and CRISPR-based genetic screens [1,2,3].
What is the role of CFTR in ATP transport?
CFTR is an ABC transporter that binds and hydrolyzes ATP to gate chloride conductance; it also transports ATP, influencing purinergic signaling [2,5].
Can CRISPR be used to model ATP transporter mutations?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are widely used to study ATP transporter gene function and disease mutations [3,6].
What is the mitochondrial permeability transition pore?
The mPTP is a high-conductance channel that opens under stress, leading to ATP depletion and cell death; ATP transporters are involved in its regulation.
How does P-glycoprotein contribute to drug resistance?
P-glycoprotein (ABCB1) uses ATP hydrolysis to efflux chemotherapeutic drugs, reducing their intracellular concentration and causing multidrug resistance.
What services does EDITGENE offer for ATP transporter research?
EDITGENE provides CRISPR knockout, point mutation, knock-in, overexpression models, library screening, and bioinformatics services tailored to ATP transporter genes.
Conclusion
ATP transmembrane transporter activity (GO:0005347) is a fundamental molecular function that governs cellular energy distribution and signaling. Its dysregulation underlies a diverse array of human diseases, from mitochondrial disorders to cancer. Advances in CRISPR technology have revolutionized the study of ATP transporters by enabling precise genetic manipulation in relevant cell models. EDITGENE stands at the forefront of this field, offering comprehensive services to help researchers uncover the mechanisms and therapeutic potential of ATP transmembrane transporter activity.
References
- 1. Bonora M et al.. 2022. Molecular mechanisms and consequences of mitochondrial permeability transition.. Nat Rev Mol Cell Biol 23(4):266-285 PMID: 34880425
- 2. Kirk KL et al.. 2011. A unified view of cystic fibrosis transmembrane conductance regulator (CFTR) gating: combining the allosterism of a ligand-gated channel with the enzymatic activity of an ATP-binding cassette (ABC) transporter.. J Biol Chem 286(15):12813-9 PMID: 21296873
- 3. Bi Y et al.. 2023. Transporter-mediated Natural Product-Drug Interactions.. Planta Med 89(2):119-133 PMID: 35304735
- 4. Kaplan JH. 2002. Biochemistry of Na,K-ATPase.. Annu Rev Biochem 71:511-35 PMID: 12045105
- 5. Gross CH et al.. 2006. Nucleotide-binding domains of cystic fibrosis transmembrane conductance regulator, an ABC transporter, catalyze adenylate kinase activity but not ATP hydrolysis.. J Biol Chem 281(7):4058-68 PMID: 16361259
- 6. Becker PC et al.. 2024. Identification of Human TRIAC Transmembrane Transporters.. Thyroid 34(7):920-930 PMID: 38801167
- 7. Pan Y et al.. 2026. Increased ATP production and P-glycoprotein activity underlie the marked changes in blood-brain barrier transport of drugs in a mouse model of amyotrophic lateral sclerosis.. Br J Pharmacol 183(2):280-295 PMID: 40891024
- 8. Zolnerciks JK et al.. 2011. Structure of ABC transporters.. Essays Biochem 50(1):43-61 PMID: 21967051
