Electron-capture dissociation

Electron-capture dissociation (ECD) is a method of fragmentation used in mass spectrometry to analyze highly complex molecules, such as proteins, peptides, and other biopolymers. Unlike other fragmentation techniques, ECD involves the direct capture of low-energy electrons by multiply-charged molecular ions. This process results in cleavage of the N-Cα bond of the peptide backbone while largely preserving the more labile post-translational modifications, making ECD particularly useful in the study of protein structure and post-translational modifications.

Overview

Electron-capture dissociation was first introduced in 1998 by Roman Zubarev and Neil Kelleher. The technique is especially advantageous for the structural elucidation of biomolecules, as it allows for the differentiation of isomeric compounds and the detailed mapping of protein folding and protein-protein interactions. ECD is performed in Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers, although it has also been adapted to other types of mass spectrometers, such as Orbitrap and quadrupole time-of-flight (Q-TOF) instruments.

Mechanism

In electron-capture dissociation, multiply-charged positive ions (typically generated by electrospray ionization (ESI) or matrix-assisted laser desorption/ionization (MALDI)) are reacted with low-energy electrons. The capture of an electron by a positively charged ion leads to the formation of a radical ion, which subsequently undergoes fragmentation. The primary fragmentation pathway in ECD involves the cleavage of the N-Cα bond of the peptide backbone, resulting in c and z• ion series, according to the nomenclature introduced by Roepstorff and Fohlman in 1984.

Applications

ECD has found widespread application in the field of proteomics, where it is used to identify proteins and to characterize their post-translational modifications. It is particularly useful for the analysis of phosphorylation, glycosylation, and other modifications that are sensitive to the more energetic fragmentation methods. ECD is also employed in the study of protein folding and conformational changes, as it allows for the mapping of non-covalent interactions within the protein structure.

Advantages and Limitations

The main advantage of ECD is its ability to preserve labile post-translational modifications during fragmentation, providing unique insights into protein structure and function. However, the technique requires the use of specialized equipment, such as FT-ICR mass spectrometers, which can be a limitation for some laboratories. Additionally, ECD is most effective for the analysis of highly charged ions, which may limit its applicability to certain types of samples.

Comparison with Other Techniques

ECD is often compared with other fragmentation techniques such as collision-induced dissociation (CID) and electron-transfer dissociation (ETD). While CID is more widely available and applicable to a broader range of molecules, it often leads to the loss of labile post-translational modifications. ETD, on the other hand, is similar to ECD in its ability to preserve these modifications but differs in the mechanism of electron transfer.

Conclusion

Electron-capture dissociation has significantly advanced the field of mass spectrometry-based proteomics by enabling the detailed analysis of protein structures and post-translational modifications. Despite its limitations, the unique insights provided by ECD into the structure and function of biomolecules make it an invaluable tool in the study of biological systems.

   This article is a Mass spectrometry-related stub. You can help WikiMD by expanding it!

Electron-capture dissociation

• 
  Schematic diagram of the combined ECD FTICRMS and IRMPD experimental setup
• 
  Schematic of AP-ECD source