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Related Concept Videos

ATP Synthase: Structure01:18

ATP Synthase: Structure

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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...
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ATP Synthase: Mechanism01:48

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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...
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Overview of Nitrogen Metabolism01:20

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Nitrogen is a very important element for life because it is a major constituent of proteins and nucleic acids. It is a macronutrient, and in nature, it is recycled from organic compounds and stored in the form of  ammonia, ammonium ions, nitrate, nitrite, or  nitrogen gas by many metabolic processes. Many of these metabolic processes are carried out only by prokaryotes.
The largest pool of nitrogen available in the terrestrial ecosystem is gaseous nitrogen (N2) from the air, but this...
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ATP Energy Storage and Release01:31

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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.
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ATP Driven Pumps I: An Overview01:27

ATP Driven Pumps I: An Overview

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ATP-driven pumps, also known as transport ATPases, are integral membrane proteins. They have binding sites for ATP located on the membrane's cytosolic side and the ion-conducting domain in the transmembrane region. These pumps use the free energy released from ATP hydrolysis to move the solutes across cell membranes against an electrochemical gradient.
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Allosteric Proteins-ATCase01:19

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Binding sites linkages can regulate a protein's function.  For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
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Updated: Jun 24, 2025

A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors
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A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors

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Ancient nitrogenases are ATP dependent.

Derek F Harris1, Holly R Rucker2, Amanda K Garcia2

  • 1Department of Chemistry and Biochemistry, Utah State University, Logan, Utah, USA.

Mbio
|June 13, 2024
PubMed
Summary
This summary is machine-generated.

Ancient nitrogenase enzymes strictly required adenosine triphosphate (ATP) for function, similar to modern enzymes. This research reconstructs an ancient enzyme to show ATP

Keywords:
ancestral sequence reconstructionenergynitrogen fixation

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Area of Science:

  • Biochemistry
  • Evolutionary Biology
  • Enzymology

Background:

  • Life relies on energy currencies like adenosine triphosphate (ATP) for cellular processes.
  • The evolutionary origins and ancient enzyme dependence on ATP remain unclear.
  • Reconstructing ancestral enzymes offers insights into early biochemistry.

Purpose of the Study:

  • To investigate the evolutionary necessity of ATP for ancient enzymes.
  • To experimentally reconstruct a Proterozoic-era nitrogenase ancestor.
  • To determine the nucleotide and metal ion specificity of the ancestral enzyme.

Main Methods:

  • Experimental reconstruction of an ancestral nitrogenase enzyme.
  • Assays to measure enzyme activity with various nucleotide triphosphates and divalent metal ions.
  • Comparison of ancestral enzyme activity and efficiency with the extant enzyme.

Main Results:

  • The reconstructed ancestral nitrogenase strictly required ATP, showing no activity with GTP, ITP, or UTP.
  • Magnesium (Mg2+) was the preferred divalent metal ion, though others supported reduced activity.
  • The ancestral enzyme exhibited an ATP hydrolysis efficiency of two molecules per electron transferred, matching the extant enzyme.

Conclusions:

  • Provides direct experimental evidence for ATP usage by an ancient enzyme.
  • Demonstrates a conserved, strict requirement for ATP in nitrogenase function across evolutionary time.
  • Highlights the early establishment of ATP as a critical energy currency in biological systems.