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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

ATP Synthase: Mechanism

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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 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.
There are four main types of ATP-driven pumps - P-type, V-type, F-type, and ABC transporter. All these pumps are of varying complexities and...
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ATP Driven Pumps II: P-type Pumps01:34

ATP Driven Pumps II: P-type Pumps

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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 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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Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.3K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
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Related Experiment Video

Updated: Sep 20, 2025

Visualization of ATP Synthase Dimers in Mitochondria by Electron Cryo-tomography
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Visualizing Single V-ATPase Rotation Using Janus Nanoparticles.

Akihiro Otomo1,2, Jared Wiemann3, Swagata Bhattacharyya3

  • 1Institute for Molecular Science, National Institutes of National Sciences, Okazaki, Aichi 444-8787, Japan.

Nano Letters
|November 22, 2024
PubMed
Summary
This summary is machine-generated.

This study introduces Janus nanoparticles for directly visualizing single V-ATPase motor rotation. This novel method accurately measures torque, advancing single-molecule analysis of rotary motors.

Keywords:
Janus nanoparticlesfluctuation theoremrotary ATPasesrotational trackingsingle-molecule analysistorque measurement

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Area of Science:

  • Biophysics
  • Nanotechnology
  • Biochemistry

Background:

  • Rotary molecular motors, like V-ATPases, are crucial for cellular functions.
  • Visualizing single-molecule rotation is key to understanding their mechanisms.
  • Conventional nanoparticle (NP) tracking methods infer rotation indirectly.

Purpose of the Study:

  • To develop a direct imaging method for single V-ATPase rotation using Janus NPs.
  • To assess the accuracy of Janus NPs in torque measurements for rotary motors.
  • To highlight the advantages of Janus NPs over traditional probes.

Main Methods:

  • Utilized silica/gold Janus NPs with asymmetric optical contrast for imaging.
  • Immobilized single V-ATPase motors from *Enterococcus hirae* on surfaces.
  • Analyzed the unidirectional counterclockwise rotation and measured torque.

Main Results:

  • Successfully imaged the direct rotation of single V-ATPase motors.
  • Demonstrated accurate torque measurements despite the viscous load of the Janus NP.
  • Showcased the superior performance of Janus NPs compared to conventional probes.

Conclusions:

  • Janus NPs provide a powerful tool for direct single-molecule visualization of rotary motors.
  • This approach enhances our understanding of V-ATPase function and torque generation.
  • The method has broad applicability for studying other rotary molecular machines.