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Related Concept Videos

Hydrolysis of ATP01:08

Hydrolysis of ATP

The bonds of adenosine triphosphate (ATP) can be broken through the addition of water, releasing one or two phosphate groups in an exergonic process called hydrolysis. This reaction liberates the energy in the bonds for use in the cell—for instance, to synthesize proteins from amino acids.
If one phosphate group is removed, a molecule of ADP—adenosine diphosphate—remains, along with inorganic phosphate. ADP can be further hydrolyzed to AMP—adenosine monophosphate—by the removal of a second...
ATP Synthase: Mechanism01:48

ATP Synthase: Mechanism

In animals, the mitochondrial F1F0 ATP synthase is the key protein that synthesizes ATP molecules through a complex catalytic mechanism. While the nuclear genome encodes the majority of ATP synthase subunits, the mitochondrial genome encodes some of the enzyme's most critical components. The formation of this multi-subunit enzyme is a complex multi-step process regulated at the level of transcription, translation, and assembly. Defects in one or more of these steps can result in decreased ATP...
ATP Synthase: Structure01:18

ATP Synthase: Structure

ATP synthase or ATPase is among the most conserved proteins found in bacteria, mammals, and plants. This enzyme can catalyze a forward reaction in response to the electrochemical gradient, producing ATP from ADP and inorganic phosphate. ATP synthase can also work in a reverse direction by hydrolyzing ATP and generating an electrochemical gradient. Different forms of ATP synthases have evolved special features to meet the specific demands of the cell. Based on their specific feature, ATP...
ATP Energy Storage and Release01:31

ATP Energy Storage and Release

ATP is a highly unstable molecule. Unless quickly used to perform work, ATP spontaneously dissociates into ADP and inorganic phosphate (Pi), and the free energy released during this process is lost as heat. The energy released by ATP hydrolysis is used to perform work inside the cell and depends on a strategy called energy coupling. Cells couple the exergonic reaction of ATP hydrolysis with endergonic reactions, allowing them to proceed.
One example of energy coupling using ATP involves a...
Stringent Response in E. coli01:23

Stringent Response in E. coli

Bacterial growth is closely tied to nutrient availability, with cells proliferating exponentially under favorable conditions and entering a stationary phase when resources become scarce. This transition is mediated by a regulatory mechanism known as the stringent response, which allows bacteria to adapt to nutrient deprivation by modulating gene expression and metabolic activity.During nutrient scarcity, intracellular amino acid levels decline. It results in the accumulation of uncharged tRNAs...

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Related Experiment Video

Updated: Jul 6, 2026

Direct Detection of the Acetate-forming Activity of the Enzyme Acetate Kinase
05:51

Direct Detection of the Acetate-forming Activity of the Enzyme Acetate Kinase

Published on: December 19, 2011

Kinetic properties of soluble adenosine triphosphatase of Escherichia coli.

J Ahlers

    Molecular and Cellular Biochemistry
    |April 12, 1977
    PubMed
    Summary

    ATPase from Escherichia coli exhibits similar kinetic properties whether bound or solubilized. MgATP acts as the true substrate, with no cooperativity observed in enzyme binding.

    Area of Science:

    • Biochemistry
    • Enzymology
    • Molecular Biology

    Background:

    • ATPase enzymes are crucial for cellular energy metabolism.
    • Understanding ATPase kinetics is essential for elucidating enzyme mechanisms.
    • Escherichia coli ATPase serves as a model system for studying enzyme function.

    Purpose of the Study:

    • To compare the kinetic properties of bound and solubilized ATPase from Escherichia coli.
    • To determine the substrate binding and cooperativity of MgATP with ATPase.
    • To investigate the effect of chelating agents on ATPase kinetics.

    Main Methods:

    • Enzyme kinetic assays were performed on both bound and solubilized ATPase preparations.
    • Saturation curves for MgATP were analyzed using hyperbolic and sigmoidal models.

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    Last Updated: Jul 6, 2026

    Direct Detection of the Acetate-forming Activity of the Enzyme Acetate Kinase
    05:51

    Direct Detection of the Acetate-forming Activity of the Enzyme Acetate Kinase

    Published on: December 19, 2011

    Measuring In Vitro ATPase Activity for Enzymatic Characterization
    07:38

    Measuring In Vitro ATPase Activity for Enzymatic Characterization

    Published on: August 23, 2016

    Utilizing Thermal Shift Assay to Probe Substrate Binding to Selenoprotein O
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  • Line-weaver-Burk plots were utilized to assess substrate binding and cooperativity.
  • The influence of ethylenediaminetetraacetic acid (EDTA) and magnesium chloride (MgCl2) on enzyme kinetics was examined.

    • Main Results:

    • Both bound and solubilized ATPase preparations displayed similar hyperbolic MgATP saturation curves.
    • Line-weaver-Burk analysis confirmed MgATP as the true substrate, with a 1:1 enzyme-substrate stoichiometry and no cooperativity.
    • The presence of EDTA induced sigmoidal saturation curves, indicative of altered binding.
    • Stoichiometric addition of MgCl2 reversed the EDTA-induced sigmoidal effect, restoring hyperbolic kinetics.

    Conclusions:

    • MgATP is the true substrate for Escherichia coli ATPase, binding with a 1:1 stoichiometry and no cooperativity.
    • Chelating agents like EDTA can significantly alter ATPase kinetics, leading to apparent sigmoidal behavior.
    • Observed discrepancies in previous studies may be attributed to contaminations with complexing agents in assay components.