GO:0016887 ATP hydrolysis activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0016887 ATP hydrolysis activity describes the catalysis of ATP + H2O = ADP + H+ phosphate, a fundamental reaction that provides energy for many cellular processes.
• ATP hydrolysis is required for diverse functions including proteolysis, DNA mismatch repair, chromatin remodeling, and antiviral signaling.
• Key proteins with ATP hydrolysis activity include chaperones, proteases, helicases, and ATPases such as HslU, MutL, ISWI, and CRISPR-associated proteins.
• Dysregulation of ATP hydrolysis is linked to diseases such as cancer, fibrosis, and viral infections.
• Research methods to study ATP hydrolysis include radiolabeled ATP assays, structural biology, and CRISPR-based gene editing.
• CRISPR knockout, point mutation, knock-in, and overexpression models enable precise interrogation of ATP hydrolysis genes in disease and development.
Description
ATP hydrolysis activity (GO:0016887) is a molecular function that catalyzes the conversion of ATP and water into ADP, inorganic phosphate, and a proton, releasing free energy that drives numerous cellular processes. This activity is essential for energy transduction in cells, powering reactions such as protein degradation, DNA repair, and chromatin remodeling. Researchers study ATP hydrolysis to understand how cells utilize energy and how defects in this process contribute to disease. The reaction is fundamental to life, and its precise regulation ensures proper cellular function.
ATP hydrolysis activity At A Glance
| GO ID | GO:0016887 |
|---|---|
| GO term | ATP hydrolysis activity |
| Ontology | molecular_function |
| Synonym | adenosine 5'-triphosphatase activity, adenosine triphosphatase activity, adenosinetriphosphatase activity, ATP hydrolase activity, ATP monophosphatase activity, ATP phosphohydrolase activity |
| Major function | Catalysis of ATP hydrolysis to ADP and phosphate, providing energy for cellular processes |
| Reaction | ATP + H2O = ADP + H+ phosphate |
| Energy coupling | Used as an energy source to drive other reactions or transport |
| Examples | Proteases, helicases, chaperones, chromatin remodelers, CRISPR-associated proteins |
What Is GO:0016887?
According to the Gene Ontology, GO:0016887 ATP hydrolysis activity is defined as the catalysis of the reaction: ATP + H2O = ADP + H+ phosphate. This activity is used in some reactions as an energy source, for example to catalyze a reaction or drive transport against a concentration gradient. It is a molecular function that encompasses enzymes known as ATPases or ATP hydrolases, which break down ATP to release energy.
Why Is ATP hydrolysis activity Important in Cell Biology?
ATP hydrolysis activity is central to cellular energy metabolism and is involved in nearly every aspect of cell biology, from protein quality control to gene regulation. Its dysregulation has been implicated in a wide range of diseases, including cancer, fibrosis, and viral infections. Understanding the mechanisms and regulation of ATP hydrolysis is therefore critical for developing therapeutic strategies and for basic research in molecular biology.
• Provides energy for proteolysis by the HslVU protease in bacteria.
• Required for DNA mismatch repair by MutL in an ATP-hydrolysis-dependent manner.
• Drives chromatin remodeling by ISWI, affecting gene expression.
• Enables antiviral signaling by type III CRISPR-associated deaminases.
• Regulates fibrotic activity of cardiac fibroblasts through extracellular ATP hydrolysis.
• Supports geminivirus replication by interfering with ATP hydrolysis of replication initiator protein.
• Used as a detection principle for alkaline phosphatase activity via CRISPR/Cas12a.
• Measured using radiolabeled ATP assays for enzyme kinetics.
• Involved in many ATPases that are targets for drug discovery.
• Essential for cellular homeostasis and energy balance.
What Happens During ATP hydrolysis activity?
Substrate Binding and Activation
In simple terms: The enzyme grabs ATP and water, getting ready to break ATP apart.
ATP hydrolysis begins with the binding of ATP and a water molecule to the active site of the enzyme. This binding often induces conformational changes that position the ATP for cleavage. For example, in the HslVU protease, ATP binding, but not its hydrolysis, is required for assembly and proteolytic activity, indicating that nucleotide binding is a key regulatory step. Similarly, MutL requires ATP hydrolysis to stimulate the endonuclease activity of MutS in DNA mismatch repair.
Catalysis and Product Release
In simple terms: The enzyme splits ATP into ADP and phosphate, releasing energy.
The catalytic step involves the nucleophilic attack of water on the gamma-phosphate of ATP, resulting in the formation of ADP and inorganic phosphate. This reaction releases free energy that can be used to drive other processes. For instance, ISWI chromatin remodeler uses ATP hydrolysis to slide nucleosomes and regulate gene expression. The products, ADP and phosphate, are then released from the active site, allowing the enzyme to cycle again.
Energy Coupling to Cellular Processes
In simple terms: The energy from ATP breakdown powers other jobs in the cell.
The energy released by ATP hydrolysis is coupled to various cellular activities. In cardiac fibroblasts, hydrolysis of extracellular ATP by ENTPD establishes a set point for fibrotic activity, linking ATP hydrolysis to tissue remodeling. In antiviral signaling, type III CRISPR-associated deaminases require ATP hydrolysis to activate their antiviral response. Additionally, geminivirus-encoded replication initiator protein's ATP hydrolysis activity is targeted by betasatellite-encoded βC1 protein to regulate viral accumulation.
Regulation and Feedback
In simple terms: The cell controls when and where ATP is broken down.
ATP hydrolysis is tightly regulated to match cellular energy demands. For example, the activity of MutL is stimulated by MutS in an ATP-hydrolysis-dependent manner, ensuring mismatch repair occurs only when needed. In fibrotic cardiac fibroblasts, extracellular ATP hydrolysis sets a threshold for fibrotic activity, indicating a regulatory role. Furthermore, ATP hydrolysis by ISWI is modulated during chromatin remodeling to maintain proper gene expression.
Key Genes Involved in GO:0016887 ATP hydrolysis activity
The following genes and proteins are key players in ATP hydrolysis activity, as evidenced by published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| HslU | ATP-dependent protease in E. coli | Studied for ATP binding vs hydrolysis in proteolysis |
| MutL | DNA mismatch repair | ATP hydrolysis stimulates MutS endonuclease |
| ISWI | Chromatin remodeling | Structural insights into ATP hydrolysis during nucleosome sliding |
| ENTPD | Extracellular ATP hydrolysis | Regulates fibrotic activity in cardiac fibroblasts |
| βC1 | Viral protein interfering with ATP hydrolysis | Regulates geminivirus replication |
| CRISPR-associated deaminase | Antiviral signaling | Requires ATP hydrolysis for activation |
| Alkaline phosphatase | ATP hydrolysis in detection assays | Used in CRISPR/Cas12a biosensing |
| ATPase (generic) | ATP hydrolysis | Measured using radiolabeled ATP |
| HslV | Protease component | Assembly requires ATP binding |
| MutS | Mismatch recognition | Stimulates MutL endonuclease via ATP hydrolysis |
| Cas12a | CRISPR effector | Used in ATP hydrolysis-based detection |
| Replication initiator protein | Viral replication | ATP hydrolysis activity targeted by βC1 |
| Ectonucleoside triphosphate diphosphohydrolase | ATP hydrolysis | Sets fibrotic set point |
| Type III CRISPR deaminase | Antiviral defense | ATP hydrolysis-dependent signaling |
| ISWI complex | Chromatin remodeling | ATP hydrolysis drives nucleosome sliding |
| HslVU protease | Protein degradation | ATP binding required for assembly |
| MutL-MutS complex | DNA repair | ATP hydrolysis-dependent activation |
How Is ATP hydrolysis activity Regulated?
ATP hydrolysis activity is regulated at multiple levels, including substrate availability, post-translational modifications, and interaction with regulatory proteins. For example, MutL activity is stimulated by MutS in an ATP-hydrolysis-dependent manner, ensuring mismatch repair is activated only in the presence of DNA lesions. In cardiac fibroblasts, extracellular ATP hydrolysis by ENTPD establishes a set point for fibrotic activity, indicating that the level of ATP hydrolysis can modulate disease progression. Additionally, the βC1 protein of betasatellites interferes with the ATP hydrolysis activity of geminivirus replication initiator protein, thereby regulating viral accumulation.
ATP hydrolysis activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| ENTPD | Cardiac fibrosis | Knockout or overexpression in cardiac fibroblasts |
| βC1 | Geminivirus infection | Point mutation to disrupt ATP hydrolysis interference |
| MutL | Cancer (mismatch repair deficiency) | Knock-in of ATP hydrolysis-deficient mutant |
| CRISPR deaminase | Viral infection | Knockout to study antiviral signaling |
| ISWI | Chromatin remodeling disorders | Point mutation in ATPase domain |
ATP Hydrolysis in Fibrosis
Hydrolysis of extracellular ATP by ectonucleoside triphosphate diphosphohydrolase (ENTPD) establishes the set point for fibrotic activity of cardiac fibroblasts. This suggests that ATP hydrolysis plays a critical role in the development of cardiac fibrosis, and modulating this activity could be a therapeutic strategy.
ATP Hydrolysis in Viral Infections
Geminivirus-encoded replication initiator protein requires ATP hydrolysis for viral replication. The betasatellite-encoded βC1 protein interferes with this activity to regulate helper virus accumulation, highlighting a viral strategy to manipulate host ATP hydrolysis for efficient infection. Additionally, type III CRISPR-associated deaminases require ATP hydrolysis for antiviral signaling, linking ATP hydrolysis to innate immunity.
ATP Hydrolysis in Cancer and DNA Repair
MutL, a key player in DNA mismatch repair, requires ATP hydrolysis to stimulate the endonuclease activity of MutS. Defects in this process can lead to microsatellite instability and cancer predisposition, underscoring the importance of ATP hydrolysis in maintaining genomic stability.
From ATP hydrolysis activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does ATP hydrolysis by HslU drive proteolysis? | Knockout of hslU in E. coli |
| How does MutL ATP hydrolysis affect mismatch repair? | Point mutation in ATPase domain of MutL |
| What is the role of ENTPD in fibrosis? | Overexpression of ENTPD in cardiac fibroblasts |
| How does βC1 interfere with viral ATP hydrolysis? | Knock-in of βC1 into geminivirus genome |
| Does ISWI ATP hydrolysis regulate chromatin remodeling? | Tagged knock-in of ISWI for live imaging |
| Can CRISPR deaminase ATP hydrolysis be targeted? | Knockout of CRISPR deaminase in antiviral assays |
How to Study the ATP hydrolysis activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Radiolabeled ATP assay | ATP hydrolysis rate | Enzyme kinetics |
| Cryo-EM | Structural changes during ATP hydrolysis | Chromatin remodeler studies |
| CRISPR/Cas12a detection | ATP hydrolysis-coupled signal | Biosensing |
| Knockout | Loss of gene function | Protease assembly studies |
| Point mutation | Specific residue function | ATP hydrolysis-deficient mutants |
| Overexpression | Gain of function | Fibrosis models |
| Knock-in | Tagged protein expression | Live imaging |
Radiolabeled ATP Assays
ATPase activity can be measured using radiolabeled ATP, which allows sensitive detection of hydrolysis products. This method is widely used to quantify enzyme kinetics and is applicable to purified proteins or cell lysates.
Structural Biology
Structural insights into ATP hydrolysis can be obtained using X-ray crystallography or cryo-EM. For example, the ISWI chromatin remodeler was studied during active ATP hydrolysis to reveal conformational changes.
CRISPR-Based Detection
ATP hydrolysis can be coupled to CRISPR/Cas12a-based detection systems. For instance, alkaline phosphatase activity was detected based on ATP hydrolysis and CRISPR/Cas12a, demonstrating a novel biosensing application.
Genetic Knockout and Mutagenesis
Gene knockout and point mutations are powerful methods to study the role of ATP hydrolysis in vivo. For example, knockout of hslU in E. coli revealed that ATP binding, but not hydrolysis, is required for HslVU protease assembly.
How CRISPR Can Be Used to Study GO:0016887 ATP hydrolysis activity
Knockout
CRISPR knockout is used to completely abolish the expression of genes involved in ATP hydrolysis, such as hslU or ENTPD, to study their loss-of-function phenotypes. For example, knockout of hslU in E. coli demonstrated that ATP binding, but not hydrolysis, is required for HslVU protease assembly.
Point Mutation
Point mutations can be introduced to specifically disrupt ATP hydrolysis activity without affecting other functions. For instance, mutations in the ATPase domain of MutL have been used to show that ATP hydrolysis is required for stimulating MutS endonuclease activity.
Knock-in
Knock-in of tagged or mutant versions of ATP hydrolysis genes allows for precise tracking and functional studies. For example, knock-in of a tagged ISWI can be used to visualize chromatin remodeling in live cells.
Overexpression
Overexpression of ATP hydrolysis-related genes, such as ENTPD, can be used to study gain-of-function effects in disease models like cardiac fibrosis.
How EDITGENE Supports ATP hydrolysis activity Research
Researchers studying ATP hydrolysis activity-related genes often need to determine whether a candidate gene is causally involved in a specific cellular process or disease. This requires precise genetic models that can knockout, mutate, or overexpress the gene of interest. EDITGENE provides a comprehensive suite of CRISPR services to enable such studies.
Contact EDITGENE today to design your custom CRISPR model for ATP hydrolysis activity research.
Frequently Asked Questions About ATP hydrolysis activity
What is ATP hydrolysis activity?
ATP hydrolysis activity (GO:0016887) is the catalysis of ATP and water to ADP and phosphate, releasing energy for cellular processes.
What genes are involved in ATP hydrolysis activity?
Genes such as HslU, MutL, ISWI, ENTPD, and CRISPR-associated deaminases encode proteins with ATP hydrolysis activity.
How is ATP hydrolysis measured?
ATP hydrolysis can be measured using radiolabeled ATP assays, structural biology, or CRISPR-based detection systems.
Why is ATP hydrolysis important?
It provides energy for proteolysis, DNA repair, chromatin remodeling, and antiviral signaling, and its dysregulation is linked to diseases like fibrosis and cancer.
What diseases are associated with ATP hydrolysis?
Cardiac fibrosis, viral infections, and cancer have been linked to altered ATP hydrolysis activity.
Can CRISPR be used to study ATP hydrolysis?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are powerful tools to study ATP hydrolysis genes.
What is the role of ATP hydrolysis in chromatin remodeling?
ATP hydrolysis by ISWI drives nucleosome sliding, which regulates gene expression.
How does ATP hydrolysis affect viral replication?
Geminivirus replication initiator protein requires ATP hydrolysis, and viral proteins like βC1 can interfere with this activity to modulate infection.
What is the connection between ATP hydrolysis and fibrosis?
Extracellular ATP hydrolysis by ENTPD sets the set point for fibrotic activity in cardiac fibroblasts.
What methods are used to study ATP hydrolysis in cells?
Methods include radiolabeled ATP assays, CRISPR screens, structural biology, and live-cell imaging.
Conclusion
ATP hydrolysis activity (GO:0016887) is a fundamental molecular function that powers diverse cellular processes, from protein degradation to antiviral defense. Its dysregulation contributes to diseases such as fibrosis, cancer, and viral infections. Continued research using advanced CRISPR models and biochemical assays will further illuminate its mechanisms and therapeutic potential.
References
- 1. Guo Y et al.. 2026. Detection of alkaline phosphatase activity based on ATP hydrolysis and CRISPR/Cas12a.. Anal Bioanal Chem 418(8):2337-2344 PMID: 41711853
- 2. Yoo SJ et al.. 1997. ATP binding, but not its hydrolysis, is required for assembly and proteolytic activity of the HslVU protease in Escherichia coli.. Biochem Biophys Res Commun 238(2):581-5 PMID: 9299555
- 3. Shimada A et al.. 2013. MutS stimulates the endonuclease activity of MutL in an ATP-hydrolysis-dependent manner.. FEBS J 280(14):3467-79 PMID: 23679952
- 4. Swarts HG et al.. 2016. ATPase Activity Measurements Using Radiolabeled ATP.. Methods Mol Biol 1377:121-6 PMID: 26695028
- 5. Lu D et al.. 2013. Hydrolysis of extracellular ATP by ectonucleoside triphosphate diphosphohydrolase (ENTPD) establishes the set point for fibrotic activity of cardiac fibroblasts.. J Biol Chem 288(26):19040-9 PMID: 23677997
- 6. Gnanasekaran P et al.. 2023. Betasatellite-encoded βC1 protein regulates helper virus accumulation by interfering with the ATP hydrolysis activity of geminivirus-encoded replication initiator protein.. J Gen Virol 104(6) PMID: 37326617
- 7. Sia Y et al.. 2025. Structural insights into chromatin remodeling by ISWI during active ATP hydrolysis.. Science 388(6751):eadu5654 PMID: 40179160
- 8. Li Y et al.. 2025. Antiviral signaling of a type III CRISPR-associated deaminase.. Science 387(6736):eadr0393 PMID: 39666823
