Native top-down mass spectrometry for higher-order structural characterization of proteins and complexes

Area of Science:

• Structural biology
• Biochemistry
• Analytical chemistry

Background:

• High demand for advanced analytical tools in structural biology.
• Growing interest in native mass spectrometry (nMS), especially native top-down mass spectrometry (nTDMS).
• nTDMS provides multi-layered structural information for biological assemblies.

Purpose of the Study:

• To review recent advances in nTDMS for structural research.
• To highlight the unique structural information obtained through nTDMS.
• To discuss applications, challenges, and future perspectives of nTDMS.

Main Methods:

• Overview of sample preparation and ionization for nMS.
• Description of tandem fragmentation techniques and mass analyzers for nTDMS.
• Discussion of software and analysis pipelines for nTDMS data.

Main Results:

• nTDMS provides unique structural information beyond traditional methods.
• Demonstrated broad applications in diverse biological systems.
• Detailed examples of protein, protein-ligand, antibody, membrane protein, and complex analyses.

Conclusions:

• nTDMS is a powerful tool for characterizing proteins and macromolecular complexes.
• Recent advancements have expanded its utility in structural biology.
• Future perspectives indicate continued growth and application in recombinant and biological samples.