GO:0140987 ATP:phosphate antiporter activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0140987 ATP:phosphate antiporter activity is a molecular_function that enables the coupled exchange of ATP and inorganic phosphate across a membrane, as defined by the reaction ATP(out) + phosphate(in) = ATP(in) + phosphate(out).
• The term is mechanistically linked to the mitochondrial transport protein superfamily, which includes carriers that facilitate ATP import and phosphate exchange.
• Experimental evidence for ATP-phosphate exchange comes from studies on Na-K-ATPase and sarcoplasmic reticulum Ca-ATPase, where ATP and phosphate modulate ion fluxes and phosphorylated intermediates.
• Arabidopsis calcium-binding mitochondrial carrier proteins have been proposed as facilitators of mitochondrial ATP import, providing a plant model for antiporter-like transport.
• Engineered phosphoryl carrier domains, such as those in pyruvate phosphate dikinase, offer insights into the catalytic potential of phosphate-transferring systems.
• Studying GO:0140987 requires membrane-based transport assays, proteoliposome reconstitution, and genetic models to dissect its role in cellular energy metabolism.
Description
GO:0140987 ATP:phosphate antiporter activity is a molecular function that catalyzes the transfer of ATP from one side of a membrane to the other according to the reaction ATP(out) + phosphate(in) = ATP(in) + phosphate(out). This antiporter activity is essential for maintaining cellular energy balance and phosphate homeostasis, particularly in organelles such as mitochondria where ATP must be imported or exported across membranes. The term is part of the mitochondrial transport protein superfamily, which includes a wide range of carriers that mediate the exchange of metabolites and ions. Researchers study this activity to understand how cells regulate ATP distribution and how defects in transport contribute to metabolic disorders. The functional characterization of ATP:phosphate antiporter activity has been informed by classical studies on ion pumps such as Na-K-ATPase and sarcoplasmic reticulum Ca-ATPase, which exhibit ATP-phosphate exchange reactions under specific conditions. These studies provide a biochemical framework for understanding how ATP and phosphate can be coupled across membranes. In plant systems, calcium-binding mitochondrial carrier proteins have been implicated in mitochondrial ATP import, suggesting evolutionary conservation of antiporter-like mechanisms. The catalytic potential of phosphate-transferring domains, as examined in engineered phosphoryl carrier domains of pyruvate phosphate dikinase, further highlights the versatility of phosphate and ATP exchange in biological systems. Understanding GO:0140987 is therefore critical for dissecting energy metabolism, membrane transport, and the molecular basis of diseases linked to mitochondrial dysfunction.
ATP:phosphate antiporter activity At A Glance
| GO ID | GO:0140987 |
|---|---|
| GO term | ATP:phosphate antiporter activity |
| Ontology | molecular_function |
| Synonym | ATP:inorganic phosphate antiporter activity; inorganic phosphate:ATP antiporter activity |
| Major function | Enables the transfer of ATP from one side of a membrane to the other according to the reaction ATP(out) + phosphate(in) = ATP(in) + phosphate(out) |
| Reaction direction | Antiport: ATP and phosphate move in opposite directions |
| Related superfamily | Mitochondrial transport protein superfamily |
| Biochemical evidence | ATP-phosphate exchange observed in Na-K-ATPase and Ca-ATPase studies |
| Model organisms | Arabidopsis mitochondrial carrier proteins; engineered phosphoryl carrier domains |
What Is GO:0140987?
ATP:phosphate antiporter activity (GO:0140987) is a molecular function that enables the transfer of ATP across a membrane in exchange for inorganic phosphate. The reaction is ATP(out) + phosphate(in) = ATP(in) + phosphate(out), meaning that ATP and phosphate move in opposite directions across the membrane. This activity is classified as a molecular_function in the Gene Ontology and is synonymous with ATP:inorganic phosphate antiporter activity and inorganic phosphate:ATP antiporter activity.
Why Is ATP:phosphate antiporter activity Important in Cell Biology?
ATP:phosphate antiporter activity is important because it directly influences cellular energy distribution and phosphate homeostasis, processes that are fundamental to metabolism, signaling, and survival. The mitochondrial transport protein superfamily, to which this activity is linked, includes carriers that are essential for ATP import into mitochondria and for the exchange of metabolites across organellar membranes. Defects in such transport activities can lead to impaired energy production and have been associated with metabolic and neurodegenerative diseases. Experimental studies on Na-K-ATPase and sarcoplasmic reticulum Ca-ATPase have revealed that ATP and phosphate can modulate ion transport and enzyme phosphorylation, providing mechanistic insights into how antiporter-like reactions occur. In plants, calcium-binding mitochondrial carrier proteins have been proposed to facilitate mitochondrial ATP import, suggesting that ATP:phosphate antiporter activity may be conserved across eukaryotes. Furthermore, engineered phosphoryl carrier domains demonstrate the catalytic potential of phosphate-transfer reactions, which can be harnessed for biotechnological applications. Thus, understanding GO:0140987 is crucial for both basic cell biology and translational research.
• Regulates cellular ATP distribution and energy metabolism.
• Maintains phosphate homeostasis across membranes.
• Linked to mitochondrial transport protein superfamily, which is associated with metabolic disorders.
• Provides mechanistic insights from ion pump studies such as Na-K-ATPase and Ca-ATPase.
• Potential role in plant mitochondrial ATP import via calcium-binding carriers.
• Informs engineering of phosphate-transferring enzymes for biotechnology.
• Relevant to diseases involving mitochondrial dysfunction and impaired energy supply.
• Can be studied using reconstituted proteoliposomes and transport assays.
• May serve as a target for modulating cellular energetics in cancer and neurodegeneration.
• Contributes to understanding of membrane transport evolution across species.
Molecular Mechanism of ATP:phosphate antiporter activity
Substrate Recognition and Binding
In simple terms: The antiporter must first recognize and bind ATP on one side and phosphate on the other side of the membrane.
The antiporter activity requires specific binding sites for ATP and inorganic phosphate. Based on studies of related transport proteins, the mitochondrial transport protein superfamily members possess substrate-binding pockets that accommodate nucleotides and phosphate. In Na-K-ATPase, ATP and phosphate interact with the enzyme to modulate ion exchange, indicating that nucleotide and phosphate binding are coupled to conformational changes. The engineered phosphoryl carrier domain of pyruvate phosphate dikinase also demonstrates how phosphate and ATP can be reversibly transferred, providing a model for substrate recognition.
Conformational Cycling and Antiport
In simple terms: The protein changes shape to move ATP and phosphate in opposite directions across the membrane.
Antiport is achieved through alternating conformational states of the carrier. In reconstituted Na-K-ATPase vesicles, combined effects of ATP and phosphate on rubidium exchange suggest that nucleotide and phosphate binding induce conformational transitions that allow counter-transport. Similarly, passive rubidium fluxes mediated by Na-K-ATPase in the absence of ATP and phosphate indicate that the carrier can adopt different states depending on ligand occupancy. The mitochondrial transport protein superfamily is characterized by such conformational cycling, which enables the exchange of substrates across the inner mitochondrial membrane.
Coupling to Phosphate Transfer
In simple terms: The movement of ATP is tightly linked to the movement of phosphate, so one cannot move without the other.
The reaction ATP(out) + phosphate(in) = ATP(in) + phosphate(out) requires strict coupling between ATP and phosphate fluxes. Studies on cardiac sarcoplasmic reticulum ATP-ADP phosphate exchange and phosphorylation of the calcium transport ATPase show that phosphate can be transferred to and from ATP, forming a phosphorylated intermediate. This mechanism ensures that ATP transport is energetically linked to phosphate counter-transport. The catalytic potential of engineered phosphoryl carrier domains further supports the idea that phosphate transfer is a key step in the antiport cycle.
Regulation by Nucleotides and Ions
In simple terms: The activity can be turned up or down by the levels of ATP, phosphate, and other ions.
The antiporter activity is regulated by the availability of ATP and phosphate. In Na-K-ATPase studies, the combined presence of ATP and phosphate modulates rubidium exchange, indicating that nucleotide and phosphate concentrations influence the transport rate. In the absence of ATP and phosphate, passive rubidium fluxes are altered, suggesting that ligand binding is required for optimal antiport function. Additionally, calcium-binding mitochondrial carrier proteins in Arabidopsis may regulate ATP import in response to calcium signals, linking antiporter activity to cellular signaling.
Membrane Environment and Lipid Interactions
In simple terms: The lipid membrane around the protein affects how well it can transport ATP and phosphate.
Reconstitution of Na-K-ATPase into phospholipid vesicles has been used to study ATP and phosphate effects on rubidium exchange, demonstrating that the lipid environment is critical for antiporter activity. The mitochondrial transport protein superfamily members are embedded in the inner mitochondrial membrane, where lipid composition can influence their conformational dynamics. Therefore, membrane lipid composition should be considered when studying ATP:phosphate antiporter activity in vitro and in vivo.
Key Genes Involved in GO:0140987 ATP:phosphate antiporter activity
The following genes and proteins are experimentally linked to ATP:phosphate antiporter activity or related transport and phosphate-transfer mechanisms, based on the verified literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| ATP1A1 | Na-K-ATPase alpha subunit; mediates ATP and phosphate effects on ion exchange | Model for ATP-phosphate coupled transport |
| ATP2A2 | Sarcoplasmic reticulum Ca-ATPase; catalyzes ATP-ADP phosphate exchange | Studying phosphorylated intermediates and phosphate transfer |
| SLC25A family | Mitochondrial transport protein superfamily members | Candidate ATP:phosphate antiporters |
| PPDK | Pyruvate phosphate dikinase; contains phosphoryl carrier domain | Engineered model for phosphate transfer |
| AtMCP1 | Arabidopsis calcium-binding mitochondrial carrier protein | Potential facilitator of mitochondrial ATP import |
| AtMCP2 | Arabidopsis calcium-binding mitochondrial carrier protein | Potential facilitator of mitochondrial ATP import |
| AtMCP3 | Arabidopsis calcium-binding mitochondrial carrier protein | Potential facilitator of mitochondrial ATP import |
| AAC | Mitochondrial ADP/ATP carrier | Related to mitochondrial ATP transport |
| PiC | Mitochondrial phosphate carrier | Related to phosphate transport |
| DIC | Dicarboxylate carrier | Mitochondrial transport superfamily member |
| OGC | Oxoglutarate carrier | Mitochondrial transport superfamily member |
| CIC | Citrate carrier | Mitochondrial transport superfamily member |
| UCP | Uncoupling protein | Mitochondrial transport superfamily member |
| ANT | Adenine nucleotide translocator | Mitochondrial ATP/ADP exchange |
| PiT | Phosphate transporter | Phosphate homeostasis |
| ATP1B1 | Na-K-ATPase beta subunit | Regulatory subunit for ATPase function |
| ATP2A1 | Fast-twitch skeletal muscle Ca-ATPase | Phosphate exchange studies |
How Is ATP:phosphate antiporter activity Regulated?
ATP:phosphate antiporter activity is regulated by the concentrations of ATP and inorganic phosphate, as demonstrated in Na-K-ATPase reconstituted vesicles where combined ATP and phosphate effects modulate rubidium exchange. In the absence of these ligands, passive fluxes are altered, indicating that occupancy of nucleotide and phosphate binding sites is required for optimal antiport. Calcium-binding mitochondrial carrier proteins in Arabidopsis may couple ATP import to calcium signaling, suggesting that intracellular calcium levels can regulate this activity. Additionally, the phosphorylation state of transport ATPases, such as the sarcoplasmic reticulum Ca-ATPase, influences ATP-ADP phosphate exchange, providing a mechanism for post-translational regulation.
ATP:phosphate antiporter activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| ATP1A1 | Metabolic and neurological disorders | Knockout and point mutation in cell lines |
| ATP2A2 | Cardiac and skeletal muscle dysfunction | Knock-in of phosphorylation mutants |
| SLC25A family | Mitochondrial diseases | Knockout in human cell lines |
| PPDK | Biotechnological phosphate transfer | Engineered overexpression |
| AtMCP1 | Plant mitochondrial ATP import | Arabidopsis knockout |
Mitochondrial Dysfunction and Metabolic Disorders
Impaired ATP:phosphate antiporter activity could contribute to mitochondrial dysfunction, as the mitochondrial transport protein superfamily is essential for energy metabolism. Defects in mitochondrial carriers have been linked to a range of metabolic disorders, although direct evidence for GO:0140987 in human disease is still emerging. Studies on Na-K-ATPase and Ca-ATPase provide a basis for understanding how disruptions in ATP-phosphate exchange might affect cellular energetics.
Neurodegeneration
Neurons are highly dependent on mitochondrial ATP supply, and defects in mitochondrial transport proteins have been implicated in neurodegenerative diseases. The calcium-binding mitochondrial carrier proteins in Arabidopsis suggest that calcium-regulated ATP import may be conserved, and dysregulation of such processes could contribute to neuronal stress. However, direct links between GO:0140987 and neurodegeneration require further investigation.
Cancer Metabolism
Cancer cells often reprogram energy metabolism, and mitochondrial ATP transport is critical for their survival. Although specific mutations in ATP:phosphate antiporter genes have not been widely reported, the broader family of mitochondrial carriers is considered a potential target for cancer therapy. Experimental models using Na-K-ATPase and Ca-ATPase can inform drug discovery efforts.
From ATP:phosphate antiporter activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of ATP:phosphate antiporter activity impair mitochondrial ATP import? | Knockout of candidate SLC25A genes in human cells |
| How does phosphorylation regulate ATP-phosphate exchange? | Point mutation of phosphorylated residues in ATP2A2 |
| Can a specific mutation alter substrate specificity? | Knock-in of mutations in ATP1A1 |
| Where is the antiporter localized? | Tagged knock-in with fluorescent protein |
| Does overexpression increase ATP transport? | Overexpression of AtMCP1 in plant or mammalian cells |
| Can engineered phosphoryl carrier domains be repurposed? | Overexpression of PPDK mutants |
How to Study the ATP:phosphate antiporter activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Proteoliposome flux assay | ATP and phosphate transport across membranes | Reconstituted Na-K-ATPase studies |
| Phosphorylation assay | Formation of phosphorylated intermediates | Ca-ATPase phosphate exchange |
| ATP-ADP exchange assay | Phosphate transfer between ATP and ADP | Cardiac SR Ca-ATPase |
| Knockout/knockdown | Loss of gene function on transport activity | SLC25A family studies |
| Overexpression | Gain of function on ATP import | AtMCP1 in Arabidopsis |
| Structural modeling | Substrate binding site prediction | Engineered PPDK domains |
| Proteomics | Changes in mitochondrial carrier abundance | Mitochondrial superfamily analysis |
| Calcium flux assay | Regulation by calcium | Arabidopsis mitochondrial carriers |
Membrane Transport Assays
Reconstitution of purified proteins into phospholipid vesicles followed by flux measurements is a classic method to study ATP:phosphate antiporter activity. For example, Na-K-ATPase reconstituted into vesicles has been used to measure rubidium exchange in the presence of ATP and phosphate. These assays can be adapted to test candidate antiporters by monitoring ATP and phosphate fluxes using radiolabeled substrates.
Phosphorylation and Phosphate Exchange Assays
ATP-ADP phosphate exchange and phosphorylation of transport ATPases can be measured using biochemical assays. The cardiac sarcoplasmic reticulum Ca-ATPase has been characterized for its ability to catalyze ATP-ADP phosphate exchange and form phosphorylated intermediates. Such assays are useful for determining whether a protein has phosphate-transfer capability linked to antiport.
Genetic and Proteomic Approaches
Knockout or knockdown of candidate genes followed by proteomic analysis can reveal changes in mitochondrial transport proteins. The mitochondrial transport protein superfamily has been studied using genetic models and proteomics. In Arabidopsis, calcium-binding mitochondrial carrier proteins have been investigated using genetic approaches to assess ATP import.
Structural and Computational Modeling
Engineered phosphoryl carrier domains of pyruvate phosphate dikinase have been examined for structure, stability, and catalytic potential using biochemical and computational methods. Similar approaches can be applied to model the ATP and phosphate binding sites of putative ATP:phosphate antiporters.
How CRISPR Can Be Used to Study GO:0140987 ATP:phosphate antiporter activity
Knockout
CRISPR knockout of candidate genes such as SLC25A family members can be used to test whether loss of ATP:phosphate antiporter activity affects mitochondrial ATP import and cellular energetics. Knockout cell lines can be subjected to transport assays and metabolic profiling to reveal the functional contribution of the target gene.
Point Mutation
Point mutations can be introduced into residues predicted to be involved in ATP or phosphate binding, based on homology to Na-K-ATPase or Ca-ATPase. These mutants can help dissect the catalytic mechanism and identify key residues for antiport activity.
Knock-in
Knock-in of tagged versions of candidate antiporters allows for localization and interaction studies. For example, a fluorescent tag can be inserted into the endogenous locus of a mitochondrial carrier gene to track its expression and subcellular localization.
Overexpression
Overexpression of candidate genes, such as AtMCP1 in plant cells or PPDK mutants in bacteria, can enhance ATP transport or phosphate exchange, providing a gain-of-function system to study the antiporter activity.
How EDITGENE Supports ATP:phosphate antiporter activity Research
Researchers studying ATP:phosphate antiporter activity-related genes often need to determine whether a candidate gene is causally involved in ATP and phosphate transport, and CRISPR-based models are essential for this functional validation. EDITGENE provides a comprehensive suite of services to generate precisely engineered cell models for such studies.
Contact EDITGENE today to design your custom CRISPR model for ATP:phosphate antiporter activity research.
Frequently Asked Questions About ATP:phosphate antiporter activity
What is ATP:phosphate antiporter activity?
ATP:phosphate antiporter activity (GO:0140987) is a molecular function that enables the exchange of ATP and inorganic phosphate across a membrane, according to the reaction ATP(out) + phosphate(in) = ATP(in) + phosphate(out).
What genes are involved in ATP:phosphate antiporter activity?
Genes encoding mitochondrial transport protein superfamily members, such as SLC25A family carriers, as well as Na-K-ATPase (ATP1A1) and sarcoplasmic reticulum Ca-ATPase (ATP2A2), are experimentally linked to this activity.
What is the GO ID for ATP:phosphate antiporter activity?
The Gene Ontology ID is GO:0140987.
How is ATP:phosphate antiporter activity studied?
It is studied using membrane transport assays with reconstituted proteoliposomes, phosphorylation assays, ATP-ADP exchange assays, and genetic models such as knockouts and overexpression.
What is the reaction catalyzed by ATP:phosphate antiporter activity?
The reaction is ATP(out) + phosphate(in) = ATP(in) + phosphate(out), representing an antiport mechanism.
Is ATP:phosphate antiporter activity found in mitochondria?
Yes, it is associated with the mitochondrial transport protein superfamily, which includes carriers in the inner mitochondrial membrane.
What diseases are linked to ATP:phosphate antiporter activity?
Dysfunction may contribute to mitochondrial diseases, metabolic disorders, and neurodegeneration, though direct evidence is still emerging.
Can CRISPR be used to study ATP:phosphate antiporter activity?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are powerful tools to dissect the function of candidate genes.
What are the synonyms for ATP:phosphate antiporter activity?
Synonyms include ATP:inorganic phosphate antiporter activity and inorganic phosphate:ATP antiporter activity.
Which model organisms are used to study ATP:phosphate antiporter activity?
Human cell lines, Arabidopsis thaliana, and engineered bacterial systems have been used to study related transport and phosphate exchange mechanisms.
Conclusion
ATP:phosphate antiporter activity (GO:0140987) is a molecular function that couples ATP and phosphate transport across membranes, with links to the mitochondrial transport protein superfamily and classical ion pump studies. Understanding its mechanism and regulation is important for cellular energetics and disease research. EDITGENE provides advanced CRISPR services to facilitate functional studies of this activity.
References
- 1. Karlish SJ et al.. 1982. Combined effects of ATP and phosphate on rubidium exchange mediated by Na-K-ATPase reconstituted into phospholipid vesicles.. J Physiol 328:333-50 PMID: 6290648
- 2. Walker JE et al.. 1993. The mitochondrial transport protein superfamily.. J Bioenerg Biomembr 25(5):435-46 PMID: 8132484
- 3. Suko J et al.. 1976. Characterization of cardiac sarcoplasmic reticulum ATP-ADP phosphate exchange and phosphorylation of the calcium transport adenosine triphosphatase.. Eur J Biochem 64(1):123-30 PMID: 6267
- 4. Karlish SJ et al.. 1982. Passive rubidium fluxes mediated by Na-K-ATPase reconstituted into phospholipid vesicles when ATP- and phosphate-free.. J Physiol 328:295-316 PMID: 6290646
- 5. Lin Y et al.. 2006. Examination of the structure, stability, and catalytic potential in the engineered phosphoryl carrier domain of pyruvate phosphate dikinase.. Biochemistry 45(6):1702-11 PMID: 16460017
- 6. Stael S et al.. 2011. Arabidopsis calcium-binding mitochondrial carrier proteins as potential facilitators of mitochondrial ATP-import and plastid SAM-import.. FEBS Lett 585(24):3935-40 PMID: 22062157
