ATPase and GTPase Profiling Services

CD Biosynsis offers comprehensive ATPase and GTPase Profiling Services, vital for characterizing major regulatory and motor proteins involved in signal transduction, cellular transport, and muscle contraction. These enzymes, which hydrolyze ATP or GTP to release energy, are significant therapeutic targets, especially in areas like oncology (GTPases like Ras) and infectious disease (viral ATPases). Our platform delivers precise kinetic and mechanistic characterization, including determining Km, kcat, and the influence of regulatory factors. We utilize highly sensitive, low-background detection technologies and offer flexibility for high-quality inhibitor screening (IC50, Ki) against large, customized ATPase/GTPase panels. Our services provide the robust and reliable data needed to understand enzyme function, validate therapeutic targets, and advance lead compounds through preclinical development.

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Quantifying Energy Hydrolysis and Regulation

ATPases and GTPases function as molecular switches or motors, and their accurate kinetic profiling is often challenging due to their dependence on membrane association, complex regulatory subunits, or low intrinsic activity. Our expertise lies in developing stable, reliable assay conditions that maintain the functional integrity of these targets. We provide access to validated recombinant proteins for diverse targets, from ion pumps and motor proteins (ATPases) to small GTPase regulatory cycles. This flexible service enables rapid identification and validation of inhibitors against specific targets (e.g., K-Ras, H-Ras), and determination of selectivity against related off-targets, ensuring a focused and efficient path toward lead optimization.

ATPase/GTPase Profiling Workflow

A structured process for rapid and reliable inhibitor characterization.

Target Panel Selection and Sourcing

Assay Platform Selection and Optimization

Screening and Potency Measurement

Advanced Kinetics and Reporting

Panel Customization: Finalize the list of ATPase and GTPase targets based on client selection and therapeutic focus.

Enzyme Quality Control: Verify purity (> 90 percent) and activity of all recombinant enzymes and required regulatory subunits.

Method Selection: Choose the most appropriate detection method (e.g., Malachite Green, EnzChek, or coupled assay) for each selected target.

Optimization: Adjust pH, cofactors (Mg2+), and enzyme concentrations to ensure linear kinetics and high assay quality (Z-factor > 0.6).

IC50 Profiling: Run dose-response curves for inhibitors against the custom-selected panel to calculate accurate IC50 values.

Selectivity Screening: Profile lead compounds against related targets in the panel to assess specificity.

Mechanistic Analysis: Perform full kinetic studies to determine the inhibition constant (Ki) and inhibition mode.
Regulatory Studies: If selected, assess the functional impact of GAPs/GEFs on the target GTPase activity.
Reporting: Deliver comprehensive reports including raw data, analyzed plots, kinetic parameters, and technical protocols.

Specialized Platforms for Nucleotide-Binding Enzymes

Customizable Enzyme Panel

Freedom to select and combine specific ATPase and GTPase targets for highly focused screening and profiling.

Complex Target Handling

Expertise in profiling challenging membrane-associated ATPases and multi-component GTPase signaling cycles.

Full Kinetic Detail

Accurate Km, kcat, and Ki determination for both nucleotide and inhibitor, guiding lead optimization efforts.

Low-Background Assays

Use of sensitive, low-interference detection methods (e.g., Malachite Green) for optimal data quality.

Client Testimonials on ATPase/GTPase Profiling

"The mechanistic profiling on our Ras inhibitor was flawless. The Ki data confirmed its non-competitive mechanism, which was crucial for our patent strategy."

Dr. Alan Xu, Oncology Drug Discovery

"Profiling our membrane-bound ion pump required specialized handling. CD Biosynsis successfully optimized the assay, delivering reproducible kcat values under physiologically relevant conditions."

Ms. Elena Rodriguez, Membrane Protein Research

"Their HTS screening of our 50,000-compound library against our viral ATPase was rapid and high-quality, yielding several promising low-micromolar hits for follow-up."

Mr. Ben Carter, Anti-Infective Program Lead

FAQs about ATPase and GTPase Profiling

Still have questions?

What is the primary product measured in ATPase/GTPase assays?

The primary product measured is inorganic phosphate (Pi), which is released when ATP or GTP is hydrolyzed to ADP or GDP, respectively. Colorimetric methods like Malachite Green are commonly used for detection.

How do you profile the function of small GTPases like Ras?

We characterize the intrinsic hydrolysis rate and also profile the modulatory effects of Guanine Nucleotide Exchange Factors (GEFs) and GTPase Activating Proteins (GAPs) on the overall regulatory cycle using fluorescence-based nucleotide exchange assays.

Why is magnesium (Mg2+) typically required for these assays?

Mg2+ is an essential cofactor because the actual substrate for most ATPases and GTPases is the metal-nucleotide complex (Mg-ATP or Mg-GTP), not the free nucleotide.

What is the advantage of a coupled enzyme assay?

Coupled assays link the Pi release to a downstream reaction (e.g., involving pyruvate kinase and lactate dehydrogenase) which consumes NAD(P)H. This provides a continuous, real-time readout of activity, even for enzymes that are active in challenging or colored buffers.

How much does Metabolic Engineering services cost?

The cost of Metabolic Engineering services depends on the project scope, complexity of the target compound, the host organism chosen, and the required yield optimization. We provide customized quotes after a detailed discussion of your specific research objectives.

Do your engineered strains meet regulatory standards?

We adhere to high quality control standards in all strain construction and optimization processes. While we do not handle final regulatory approval, our detailed documentation and compliance with best laboratory practices ensure your engineered strains are prepared for necessary regulatory filings (e.g., GRAS, FDA).

What to look for when selecting the best gene editing service?

We provide various gene editing services such as CRISPR-sgRNA library generation, stable transformation cell line generation, gene knockout cell line generation, and gene point mutation cell line generation. Users are free to select the type of service that suits their research.

Does gene editing allow customisability?

Yes, we offer very customised gene editing solutions such as AAV vector capsid directed evolution, mRNA vector gene delivery, library creation, promoter evolution and screening, etc.

What is the process for keeping data private and confidential?

We adhere to the data privacy policy completely, and all customer data and experimental data are kept confidential.

ATPase and GTPase Profiling Services

Quantifying Energy Hydrolysis and Regulation

ATPase and GTPase Target Profiling Options

Selectable GTPase Panel (Examples)

Choose Key GTPase Targets for Profiling

Selectable ATPase Panel (Examples)

Choose Key ATPase Targets for Profiling

Available Profiling Methods for Selected Targets

Assay Types and Kinetic Services

ATPase/GTPase Profiling Workflow

Specialized Platforms for Nucleotide-Binding Enzymes

Client Testimonials on ATPase/GTPase Profiling

FAQs about ATPase and GTPase Profiling