GO:0042626 ATPase-coupled transmembrane transporter activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0042626 describes primary active transporters that use ATP hydrolysis to move solutes across membranes, often against concentration gradients.
• The term covers P-type ATPases, ABC transporters, and other families that couple ATP hydrolysis to transmembrane transport.
• These transporters are essential for ion homeostasis, nutrient uptake, toxin efflux, and drug resistance in diverse organisms [1,2,5].
• Structural and mechanistic studies of PIB-type copper ATPases reveal transient phosphorylation and conformational changes during transport.
• Transcriptomic and proteomic analyses link expression of ATPase-coupled transporters to stress responses, including ivermectin resistance and ectoine biosynthesis [2,5].
• CRISPR-based knockout, point mutation, and knock-in models enable causal testing of transporter function in disease and physiology [1,2].
Description
ATPase-coupled transmembrane transporter activity (GO:0042626) defines a class of primary active transporters that hydrolyze ATP to drive the movement of solutes across biological membranes, frequently against their concentration gradients. This molecular function is fundamental to cellular physiology, enabling the accumulation of essential ions and nutrients, the extrusion of toxic compounds, and the maintenance of electrochemical gradients [1,5]. The term encompasses diverse protein families, including P-type ATPases, ABC transporters, and other ATP-hydrolyzing pumps, each with distinct structural architectures but a shared coupling of ATP hydrolysis to transmembrane transport. Researchers study this activity to understand membrane transport mechanisms, drug resistance, and the molecular basis of diseases caused by transporter dysfunction [1,2,5]. The availability of high-resolution structures, such as the copper-transporting PIB-type ATPase, has provided mechanistic insights into how ATP binding and phosphorylation drive conformational changes that translocate substrates. In addition, transcriptomic and proteomic approaches have revealed how expression of ATPase-coupled transporters is regulated under stress conditions, including low temperature, osmotic stress, and xenobiotic exposure [2,3,5]. These findings underscore the broad biological and biomedical relevance of GO:0042626, from microbial adaptation to human disease.
ATPase-coupled transmembrane transporter activity At A Glance
| GO ID | GO:0042626 |
|---|---|
| GO term | ATPase-coupled transmembrane transporter activity |
| Ontology | molecular_function |
| Synonym | ATPase activity, coupled to transmembrane movement of substances; ATP-dependent transmembrane transporter activity; P-P-bond-hydrolysis-driven transporter |
| Major function | Primary active transport of solutes across membranes driven by ATP hydrolysis |
| Transport mechanism | May involve transient phosphorylation (P-type) or no phosphorylated intermediate (ABC-type) |
| Directionality | Transport occurs up the solute's concentration gradient |
| Energy source | ATP hydrolysis (ATP + H2O = ADP + phosphate) |
What Is GO:0042626?
GO:0042626 describes a primary active transporter that moves a solute across a membrane using the energy released by ATP hydrolysis (ATP + H2O = ADP + phosphate). The transport protein may be transiently phosphorylated, as in P-type ATPases, or may operate without a phosphorylated intermediate, as in ABC transporters and other families. This activity drives transport up the solute's concentration gradient, meaning it can accumulate substances against their gradient using a primary energy source.
Why Is ATPase-coupled transmembrane transporter activity Important in Cell Biology?
ATPase-coupled transmembrane transporter activity is essential for maintaining cellular homeostasis, nutrient acquisition, and protection against toxic substances. It underlies the function of P-type ATPases, ABC transporters, and other pumps that are critical for ion balance, metal detoxification, and multidrug resistance [1,5]. In pathogens and pests, these transporters contribute to drug resistance, as shown by transcriptomic studies of ivermectin resistance in Haemonchus contortus. In biotechnology and environmental adaptation, ATPase-coupled transporters are involved in osmolyte accumulation and stress responses, such as ectoine biosynthesis in Halomonas campaniensis. Understanding this activity is therefore central to pharmacology, toxicology, and molecular physiology.
• Maintains ion gradients essential for nerve, muscle, and epithelial function.
• Enables nutrient uptake and metal homeostasis across membranes.
• Mediates multidrug resistance by effluxing xenobiotics and drugs.
• Contributes to anthelmintic resistance in parasitic nematodes.
• Supports microbial adaptation to osmotic and UV stress.
• Provides targets for therapeutic intervention in cancer and infectious diseases [1,5].
• Serves as a model for studying energy coupling in membrane proteins.
• Links to toxicogenomic responses to insecticides and environmental contaminants.
• Involved in low-temperature stress responses in plants.
• Relevant to amino acid composition and quality traits in aquaculture species.
What Happens During ATPase-coupled transmembrane transporter activity?
Substrate binding and ATP hydrolysis
In simple terms: The transporter grabs its target solute and uses ATP as an energy source to push it across the membrane.
The transport cycle begins with substrate binding from one side of the membrane, followed by ATP binding and hydrolysis. For P-type ATPases, ATP hydrolysis leads to transient phosphorylation of a conserved aspartate residue, which drives conformational changes that translocate the solute. In ABC transporters, ATP binding and hydrolysis occur at nucleotide-binding domains but do not involve a phosphorylated intermediate. The energy from ATP hydrolysis is used to move the solute against its concentration gradient.
Conformational changes and substrate translocation
In simple terms: The protein changes shape to carry the solute through the membrane and release it on the other side.
Structural studies of a copper-transporting PIB-type ATPase have revealed distinct conformational states that alternate between inward-facing and outward-facing orientations, allowing the substrate to be released on the opposite side of the membrane. These conformational transitions are coupled to the phosphorylation and dephosphorylation cycle, ensuring that transport is tightly linked to ATP hydrolysis. Similar mechanisms are observed in other P-type ATPases and ABC transporters, although the specific structural elements differ.
Dephosphorylation and resetting
In simple terms: The transporter removes the phosphate group and returns to its starting shape, ready for another round.
After substrate release, the transporter undergoes dephosphorylation, which resets the protein to its initial conformation. This step is essential for continuous cycling and prevents futile ATP hydrolysis. In P-type ATPases, the aspartate phosphate bond is hydrolyzed, and the protein returns to the high-affinity substrate-binding state. The overall process ensures that each ATP molecule hydrolyzed results in the transport of a defined number of solute molecules.
Regulation by cellular signals
In simple terms: Cells can adjust how active these transporters are based on their needs.
The activity of ATPase-coupled transporters can be regulated at multiple levels, including gene expression, post-translational modifications, and interaction with regulatory proteins. Transcriptomic studies have shown that expression of ABC transporters changes in response to environmental stresses such as NaCl and UV mutagenesis. In Haemonchus contortus, ivermectin resistance is associated with altered expression of transporter genes, suggesting that regulation of these pumps contributes to drug resistance. Additionally, proteomic analyses have identified lysine dihydroxyisobutyrylation as a potential regulatory modification under low-temperature stress in Dendrobium huoshanense.
Key Genes Involved in GO:0042626 ATPase-coupled transmembrane transporter activity
The following genes encode proteins that exhibit ATPase-coupled transmembrane transporter activity or are directly involved in its regulation and function.
| Gene | Major Role | Research Relevance |
|---|---|---|
| ATP7A | Copper-transporting P-type ATPase | Mutations cause Menkes disease; structural model for PIB-type ATPases |
| ATP7B | Copper-transporting P-type ATPase | Mutations cause Wilson disease; target for copper homeostasis studies |
| ABCB1 | Multidrug efflux ABC transporter | Mediates drug resistance in cancer and parasites |
| ABCC1 | Multidrug resistance-associated protein | Involved in xenobiotic efflux and toxicogenomic responses |
| ABCG2 | ABC transporter efflux pump | Affects drug bioavailability and resistance |
| ATP1A1 | Na+/K+-ATPase alpha subunit | Maintains electrochemical gradients; target of cardiac glycosides |
| ATP2A1 | SERCA calcium ATPase | Regulates calcium signaling in muscle |
| ATP2B1 | Plasma membrane calcium ATPase | Calcium homeostasis and signaling |
| ATP6V1A | Vacuolar H+-ATPase subunit | Acidifies organelles; involved in lysosomal function |
| ATP6V0A1 | Vacuolar H+-ATPase subunit | Proton transport in organelles |
| ATP8A1 | P4-type ATPase flippase | Phospholipid transport and membrane asymmetry |
| ATP10A | P4-type ATPase flippase | Lipid transport; associated with neurological disorders |
| ATP13A2 | P5-type ATPase | Mutations linked to Parkinson's disease |
| ABCB11 | Bile salt export pump | Mutations cause progressive familial intrahepatic cholestasis |
| ABCC2 | Canalicular multispecific organic anion transporter | Bile transport and drug disposition |
| ABCG5 | Sterol transporter | Cholesterol efflux; mutations cause sitosterolemia |
| ABCG8 | Sterol transporter | Cholesterol efflux; mutations cause sitosterolemia |
How Is ATPase-coupled transmembrane transporter activity Regulated?
The activity of ATPase-coupled transmembrane transporters is regulated at transcriptional, post-transcriptional, and post-translational levels. Transcriptomic analyses in Haemonchus contortus have shown that ivermectin resistance is associated with differential expression of ABC transporter genes, suggesting that upregulation of these pumps can reduce drug efficacy. In Halomonas campaniensis, expression of ABC transporters negatively correlates with ectoine biosynthesis under NaCl and UV mutagenesis treatments, indicating a trade-off between osmolyte production and transporter activity. Additionally, proteomic profiling in Dendrobium huoshanense identified lysine dihydroxyisobutyrylation as a modification that may regulate transporter function under low-temperature stress. Toxicogenomic studies in Daphnia magna revealed that exposure to fipronil and imidacloprid alters the expression of genes involved in ATPase-coupled transport, linking environmental contaminants to transporter regulation. These examples highlight the diverse regulatory mechanisms that control this activity in response to physiological and environmental cues.
ATPase-coupled transmembrane transporter activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| ATP7A | Menkes disease (copper deficiency) | Knockout or point-mutation cell models to study copper transport |
| ATP7B | Wilson disease (copper toxicity) | Knock-in of disease mutations in hepatocyte-like cells |
| ABCB1 | Multidrug resistance in cancer | Overexpression and knockout in cancer cell lines |
| ABCG2 | Drug resistance and gout | Knockout and point-mutation models in epithelial cells |
| ATP13A2 | Parkinson's disease | Knockout and knock-in models in neuronal cells |
Copper transport disorders
Mutations in the copper-transporting P-type ATPases ATP7A and ATP7B cause Menkes disease and Wilson disease, respectively. These disorders are characterized by impaired copper homeostasis, leading to severe neurological and hepatic symptoms. Structural studies of a PIB-type ATPase have provided insights into how disease-causing mutations disrupt ATP hydrolysis and copper translocation.
Multidrug resistance in cancer and infections
Overexpression of ABC transporters such as ABCB1 and ABCG2 is a major mechanism of multidrug resistance in cancer cells and pathogens. These transporters efflux chemotherapeutic agents and antimicrobials, reducing their intracellular concentrations. Transcriptomic studies in Haemonchus contortus have linked ABC transporter expression to ivermectin resistance, highlighting the clinical importance of these pumps [2,5].
Neurological and metabolic diseases
Dysfunction of ATPase-coupled transporters is associated with neurological disorders such as Parkinson's disease (ATP13A2) and metabolic conditions like sitosterolemia (ABCG5/ABCG8). These examples illustrate the broad impact of transporter dysfunction on human health [1,5].
From ATPase-coupled transmembrane transporter activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of transporter function affect ion homeostasis? | CRISPR knockout of ATP7A or ATP7B in cell lines |
| How do disease-causing point mutations alter transport activity? | Point-mutation knock-in of ATP7B variants |
| Can overexpression of ABCB1 confer drug resistance? | Overexpression of ABCB1 in cancer cell lines |
| What is the subcellular localization of a transporter? | Tagged knock-in with fluorescent protein |
| Which genes regulate transporter expression under stress? | CRISPR library screening combined with transcriptomics [2,5] |
| Does a transporter interact with regulatory proteins? | Knock-in of affinity tags for proteomics |
How to Study the ATPase-coupled transmembrane transporter activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| RNA-seq | Gene expression levels | Identifying transporters upregulated in drug resistance |
| Proteomics | Protein abundance and modifications | Detecting post-translational regulation of transporters |
| X-ray crystallography | Three-dimensional protein structure | Elucidating transport mechanism |
| ATPase activity assay | Rate of ATP hydrolysis | Measuring transporter catalytic activity |
| Transport assay | Solute flux across membranes | Validating substrate specificity and directionality |
| CRISPR knockout | Loss-of-function phenotype | Testing causal role of a transporter gene |
| CRISPR knock-in | Mutant protein expression | Modeling disease-associated mutations |
| CRISPR library screening | Pooled gene function | Identifying regulators of transporter expression [2,5] |
Transcriptomic profiling
RNA sequencing (RNA-seq) is widely used to quantify the expression of genes encoding ATPase-coupled transporters under various conditions. For example, transcriptomic analysis of Haemonchus contortus revealed differential expression of ABC transporters associated with ivermectin resistance. Similarly, RNA-seq in Halomonas campaniensis showed negative correlation between ABC transporter expression and ectoine biosynthesis under stress.
Proteomic analysis
Mass spectrometry-based proteomics can identify and quantify transporter proteins, as well as their post-translational modifications. In Dendrobium huoshanense, proteomic analysis under low-temperature stress identified lysine dihydroxyisobutyrylation as a modification potentially affecting transporter function. Proteomics also complements transcriptomics to reveal post-transcriptional regulation of transporters.
Structural biology
X-ray crystallography and cryo-electron microscopy provide high-resolution structures of ATPase-coupled transporters, revealing conformational changes during the transport cycle. The crystal structure of a copper-transporting PIB-type ATPase has elucidated the mechanism of phosphorylation-driven copper translocation.
Functional assays
ATPase activity assays measure the rate of ATP hydrolysis by transporters, often using colorimetric or fluorescent methods. Transport assays using radioisotopes or fluorescent substrates can directly measure solute translocation across membranes. These assays are essential for validating the functional impact of mutations and for screening inhibitors [1,5].
How CRISPR Can Be Used to Study GO:0042626 ATPase-coupled transmembrane transporter activity
Knockout
CRISPR knockout of genes encoding ATPase-coupled transporters enables researchers to study loss-of-function phenotypes, such as altered ion homeostasis, drug sensitivity, or substrate accumulation. For example, knocking out ATP7A or ATP7B in cell lines can reveal their roles in copper trafficking and disease pathology.
Point Mutation
Point mutations identified in patients can be introduced into endogenous genes using CRISPR base editing or homology-directed repair. This approach allows functional assessment of specific variants, such as those in ATP7B linked to Wilson disease, in an isogenic background.
Knock-in
Knock-in of reporter tags (e.g., fluorescent proteins) or affinity tags enables visualization and purification of transporters. Tagged knock-in models are valuable for studying subcellular localization, trafficking, and interaction partners of ATPase-coupled transporters.
Overexpression
CRISPR activation (CRISPRa) or lentiviral overexpression can be used to increase the expression of specific transporters, such as ABCB1, to study drug resistance mechanisms and substrate efflux capacity.
How EDITGENE Supports ATPase-coupled transmembrane transporter activity Research
Researchers studying ATPase-coupled transmembrane transporter activity-related genes often need to determine whether a candidate gene is causally involved in a specific transport process, drug resistance phenotype, or disease model. EDITGENE provides comprehensive CRISPR-based services to enable precise genetic manipulation and functional validation of transporters in relevant cell models.
Contact EDITGENE today to design your custom CRISPR model for ATPase-coupled transmembrane transporter activity research.
Frequently Asked Questions About ATPase-coupled transmembrane transporter activity
What is ATPase-coupled transmembrane transporter activity?
It is a molecular function (GO:0042626) where a protein uses ATP hydrolysis to move a solute across a membrane, often against its concentration gradient.
What genes are involved in ATPase-coupled transmembrane transporter activity?
Genes include ATP7A, ATP7B, ABCB1, ABCG2, ATP1A1, and many others encoding P-type ATPases and ABC transporters [1,5].
How does ATP hydrolysis drive transport?
ATP hydrolysis provides energy that causes conformational changes in the transporter, allowing it to translocate the substrate across the membrane.
What diseases are linked to ATPase-coupled transporters?
Mutations in ATP7A and ATP7B cause Menkes and Wilson diseases; ABC transporter overexpression causes multidrug resistance in cancer [1,5].
What is the difference between P-type and ABC transporters?
P-type ATPases form a transient phosphorylated intermediate during transport, while ABC transporters do not.
How can I study ATPase-coupled transporter activity in the lab?
Common methods include ATPase activity assays, transport assays, RNA-seq, proteomics, and CRISPR knockout models [1,2,5].
What is the role of ABC transporters in drug resistance?
ABC transporters efflux drugs out of cells, reducing intracellular drug concentrations and leading to resistance.
Can CRISPR be used to study ATPase-coupled transporters?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models enable functional studies of these transporters.
What is the GO ID for ATPase-coupled transmembrane transporter activity?
The GO ID is GO:0042626.
How are ATPase-coupled transporters regulated?
They are regulated at transcriptional, post-transcriptional, and post-translational levels, including by stress and drug exposure [2,3,5].
Conclusion
ATPase-coupled transmembrane transporter activity (GO:0042626) is a fundamental molecular function that drives primary active transport across membranes, impacting ion homeostasis, nutrient uptake, drug resistance, and disease. Structural and functional studies have elucidated the mechanisms of P-type ATPases and ABC transporters, while transcriptomic and proteomic approaches have revealed their regulation under diverse conditions [1,2,5]. CRISPR-based models provide powerful tools to dissect the causal roles of these transporters in health and disease. Continued research on GO:0042626 will advance our understanding of membrane biology and facilitate the development of targeted therapies.
References
- 1. Gourdon P et al.. 2011. Crystal structure of a copper-transporting PIB-type ATPase.. Nature 475(7354):59-64 PMID: 21716286
- 2. Reyes-Guerrero DE et al.. 2023. Assembly and Analysis of Haemonchus contortus Transcriptome as a Tool for the Knowledge of Ivermectin Resistance Mechanisms.. Pathogens 12(3) PMID: 36986421
- 3. Rao W et al.. 2025. The Role of Lysine Dihydroxyisobutyrylation in Dendrobium huoshanese Under Low-Temperature by Proteomic Analysis.. Physiol Plant 177(3):e70343 PMID: 40536206
- 5. Qiao L et al.. 2024. Expression of ABC transporters negatively correlates with ectoine biosynthesis in Halomonas campaniensis under NaCl and ultraviolet mutagenesis treatments revealed by transcriptomic and proteomics combined analysis.. BMC Genomics 25(1):1114 PMID: 39567869
- 6. Pfaff J et al.. 2021. Toxicogenomic differentiation of functional responses to fipronil and imidacloprid in Daphnia magna.. Aquat Toxicol 238:105927 PMID: 34340001
