Atp Metabolite Analysis
Atp Metabolite Analysis - Related Videos
01:31
ATP Yield
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2019
Cellular respiration produces 30 - 32 ATP per glucose molecule. Although most of the ATP results from oxidative phosphorylation and the electron transport chain (ETC), 4 ATP are gained beforehand (2 from glycolysis and 2 from the citric acid cycle). The ETC is embedded in the inner mitochondrial membrane and is comprised of four main protein complexes and an ATP synthase. NADH and FADH2 pass electrons to these complexes, which pump protons into the intermembrane space. This distribution of...
08:32
The ATP Bioluminescence Assay
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2023
In fireflies, the luciferase enzyme converts a compound called luciferin into oxyluciferin, and produces light or “luminescence” as a result. This reaction requires energy derived from ATP in order to proceed, so researchers have exploited the luciferase-luciferin interaction to gauge ATP levels in cells. Given ATP’s role as the cell’s currency of energy, the ATP bioluminescence assay can provide insight into cellular metabolism and overall cell health.In this video, JoVE discusses cellular...
11:17
A Simple Fractionated Extraction Method for the Comprehensive Analysis of Metabolites, Lipids, and Proteins from a Single Sample
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Cited by 67 •
2017
A protocol for comprehensive extraction of lipids, metabolites and proteins from biological tissues using one sample is presented.
08:03
A Tandem Liquid Chromatography–Mass Spectrometry-based Approach for Metabolite Analysis of Staphylococcus aureus
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Cited by 1 •
2017
Here we describe a protocol for the extraction of metabolites from Staphylococcus aureus and their subsequent analysis via liquid chromatography and mass spectrometry.
11:25
Multi-step Preparation Technique to Recover Multiple Metabolite Compound Classes for In-depth and Informative Metabolomic Analysis
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Cited by 29 •
2014
The reliability of results in metabolomics experiments depends on the effectiveness and reproducibility of the sample preparation. Described is a rigorous and in-depth method that enables extraction of metabolites from biological fluids with the option of subsequently analyzing up to thousands of compounds, or just the compound classes of interest.
FAQs
The relative amounts of ATP, ADP, and AMP provide more information than ATP concentration alone. Their ratios are used to indicate cellular energy charge, helping researchers determine whether cells maintain normal energy conditions or experience metabolic stress. Comparing these metabolites can therefore connect biochemical measurements with broader changes in cellular physiology.
ATP and related purine metabolites can be unstable after sample collection, so handling conditions directly affect the measurement. Rapid quenching stops or limits ongoing metabolic changes, while extraction makes the metabolites available for analysis. These preservation steps are essential because delayed processing could alter the measured relationships among ATP, ADP, and AMP.
Liquid chromatography–mass spectrometry separates metabolites before measuring them, supporting analysis of ATP and related purine compounds within the same analytical framework. Enzymatic luminescence assays use enzyme-dependent light production to quantify ATP. The overview identifies both approaches as suitable measurement strategies, allowing researchers to select an analytical format aligned with the metabolites and study design.
A typical workflow begins with rapid sample quenching, followed by metabolite extraction to preserve and recover unstable compounds. The prepared sample is then analyzed by liquid chromatography–mass spectrometry or an enzymatic luminescence assay. Researchers can compare ATP, ADP, and AMP measurements, including their ratios, to evaluate cellular energy metabolism and metabolic stress.
This analysis is useful when investigators need to connect altered energy metabolism with disease-related physiology. Supported applications include studies of mitochondrial function, ischemia, cancer metabolism, and responses to treatment. Measuring purine metabolites in these settings can help reveal metabolic defects associated with disease and support evaluation of potential therapeutic strategies.
Treatment studies can track ATP, ADP, and AMP levels or ratios before and after an intervention. Changes in these measurements may indicate altered cellular energy metabolism or metabolic stress, providing a biochemical readout of response. When interpreted alongside cellular physiology, the results can help researchers assess whether a treatment influences disease-associated metabolic defects or therapeutic mechanisms.