Related Experiment Video
Updated: Jun 9, 2026

Visualization of ATP Synthase Dimers in Mitochondria by Electron Cryo-tomography
Published on: September 14, 2014
ATP synthases: cellular nanomotors characterized by LILBID mass spectrometry
Jan Hoffmann1, Lucie Sokolova, Laura Preiss
1Institute for Physical and Theoretical Chemistry, Cluster of Excellence Frankfurt Macromolecular Complexes, Centre for Membrane Proteomics, Goethe-Universität, Max-von-Laue Str. 7, 60438 Frankfurt am Main, Germany.
This study introduces laser induced liquid bead ion desorption mass spectrometry (LILBID-MS) for analyzing membrane protein complexes like ATP synthase. The method successfully determined subunit stoichiometry in bacterial and human heart mitochondrial ATP synthases.
Area of Science:
- Biochemistry
- Molecular Biology
- Analytical Chemistry
Background:
- Mass spectrometry of membrane protein complexes presents significant challenges due to their hydrophobic/hydrophilic nature and non-covalent subunit assembly.
- Understanding the precise subunit composition of ATP synthase is crucial for elucidating its function in energy production.
Purpose of the Study:
- To apply the novel laser induced liquid bead ion desorption mass spectrometry (LILBID-MS) technique for determining the subunit stoichiometry of F(1)F(o)-ATP synthase.
- To analyze the subunit composition of ATP synthases from Bacillus pseudofirmus OF4, bovine heart, and human heart mitochondria.
Main Methods:
- Utilized laser induced liquid bead ion desorption mass spectrometry (LILBID-MS) for analyzing intact protein complexes.
- Measured the mass of intact F(1)F(o)-ATP synthase from B. pseudofirmus OF4 under specific buffer conditions.
Main Results:
- Successfully determined the subunit stoichiometry of bacterial and mitochondrial F(1)F(o)-ATP synthases.
- The measured masses of B. pseudofirmus OF4 ATP synthase closely matched theoretical masses derived from sequence databases.
- Identified differences in complexity between eukaryotic and bacterial ATP synthases, while highlighting conserved functionally important subunits.
Conclusions:
- LILBID-MS is a viable method for the mass spectrometry analysis of complex membrane protein assemblies like ATP synthase.
- Eukaryotic ATP synthases exhibit greater complexity than bacterial counterparts, yet share conserved functional subunits.
- This research provides insights into the comparative structure and composition of ATP synthases across different species.
More Related Videos
10:28A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors
Published on: August 17, 2019
10:01Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
Published on: November 28, 2017
Related Concept Videos
ATP Synthase: Structure
ATP Synthase: Mechanism
ATP Driven Pumps I: An Overview
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 are...
The ADP/ATP Carrier Protein
Chemiosmosis and ATP Synthesis
ATP Driven Pumps II: P-type Pumps
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...