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Related Concept Videos

The ADP/ATP Carrier Protein01:42

The ADP/ATP Carrier Protein

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ADP/ATP carrier or AAC protein is the most abundant carrier protein in the inner mitochondrial membrane. It transports large quantities of ADP and ATP, equivalent to the average human body weight, every day. Among other transporters, ACC protein is one of the best-studied members of the mitochondrial carrier protein family. The ADP/ATP carrier protein comprises two transmembrane helices connected to a loop and a single alpha-helix on the matrix side. It switches between two conformational...
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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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ATP Driven Pumps II: P-type Pumps01:34

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The P-type pumps are a large family of integral membrane transporter ATPases. They are divided into five major types based on substrate specificity, from I to V.
A typical P-type pump has three cytosolic domains: nucleotide-binding (N), phosphorylation (P), and activator (A) domains. These domains are connected to the membrane-spanning helices by short amino acid segments. ATP hydrolysis and covalent phosphoenzyme intermediate formation are crucial parts of the catalytic cycle. At the highly...
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ATP Synthase: Structure01:18

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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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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.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to  N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
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ATP Driven Pumps III: V-type Pumps01:30

ATP Driven Pumps III: V-type Pumps

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V-type pumps are ATP-driven pumps found in the vacuolar membranes of plants, yeast, endosomal and lysosomal membranes of animal cells, plasma membranes of a few specialized eukaryotic cells, and some prokaryotes. They are also known as the V1Vo-ATPase, that couple ATP hydrolysis to transport protons against a concentration gradient.
The peripheral or cytosolic V1 domain with eight subunits is involved in ATP hydrolysis. The integral or transmembrane V0 domain containing at least five subunits...
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Related Experiment Video

Updated: Jul 4, 2025

Measuring In Vitro ATPase Activity for Enzymatic Characterization
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Measuring In Vitro ATPase Activity for Enzymatic Characterization

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Characterizing ATP processing by the AAA+ protein p97 at the atomic level.

Mikhail Shein1,2,3, Manuel Hitzenberger4, Tat Cheung Cheng5,6

  • 1Faculty for Chemistry and Pharmacy, Ludwig-Maximilians-Universität München, München, Germany.

Nature Chemistry
|February 7, 2024
PubMed
Summary
This summary is machine-generated.

The human enzyme p97, crucial for protein homeostasis, undergoes conformational changes during ATP hydrolysis. This study reveals how p97 traps and releases phosphate, coordinating its hexameric action.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • The human enzyme p97 is vital for protein homeostasis, regulating cellular pathways by unfolding protein substrates.
  • Its hexameric structure undergoes significant conformational changes during its ATP-dependent catalytic cycle.
  • p97 is a key pharmacological target due to its essential cellular functions.

Purpose of the Study:

  • To elucidate the molecular motions at the p97 active site before and after ATP hydrolysis.
  • To understand the mechanism of ATP binding, hydrolysis, and product release in p97.
  • To reveal the coordination of hexameric subunit action and allosteric signaling.

Main Methods:

  • Integration of cryo-electron microscopy (cryo-EM) and NMR spectroscopy.
  • Application of molecular dynamics (MD) simulations.
  • Analysis of the metastable ADP·Pi reaction intermediate.

Main Results:

  • Detailed snapshots of the p97 active site trapping and discharging cleaved phosphate.
  • Identification of a metastable ADP·Pi state poised between hydrolysis and product release.
  • Elucidation of signaling pathways coordinating hexameric action and allosteric changes.

Conclusions:

  • p97's active site is finely tuned for efficient ATP hydrolysis and phosphate management.
  • Signaling pathways originating at the active site orchestrate the enzyme's complex functions.
  • This study provides insights into the spatiotemporal control of ATP hydrolysis in AAA+ proteins.