Structure-resolved virus-host interactomics by cross-linking mass spectrometry

Area of Science:

• Virology
• Structural Biology
• Biochemistry

Background:

• Viruses rely on host protein networks for replication, assembly, and spread.
• Understanding virus-host protein interactions is complex due to variability across infection stages, cell types, and species.
• Mechanistic interpretation requires detailed information on structural interfaces and conformational states.

Purpose of the Study:

• To review the advancements and applications of cross-linking mass spectrometry (XL-MS) in studying virus-host protein interactions.
• To highlight XL-MS's role in providing spatial and structural dimensions to interactomics in native systems.
• To discuss the integration of XL-MS with other techniques for a comprehensive understanding of viral mechanisms.

Main Methods:

• Cross-linking mass spectrometry (XL-MS) to map virus-host protein proximity and structure.
• Integration of XL-MS with affinity purification mass spectrometry (AP-MS).
• Complementary use of cryo-electron microscopy (cryo-EM/cryo-ET), quantitative proteomics, genetic perturbation, and structure prediction.

Main Results:

• XL-MS has evolved from analyzing viral protein complexes to mapping virion architecture and infected-cell interactomes with structural resolution.
• XL-MS effectively connects physical proximity data with molecular mechanisms of viral infection.
• The technique offers a powerful approach to study virus-host interactions in native biological systems.

Conclusions:

• XL-MS is a key technology for advancing structural virology and understanding viral pathogenesis.
• Future quantitative and integrative XL-MS workflows hold promise for systems-level structural virology.
• Addressing limitations in sensitivity, chemical coverage, and temporal resolution will further enhance XL-MS capabilities.