Methods for characterizing protein acetylation during viral infection.

Area of Science:

• Biochemistry and Molecular Biology
• Virology
• Posttranslational Modifications

Background:

• Lysine acetylation is a crucial posttranslational modification regulating protein function and interactions.
• Altered protein acetylation patterns are observed during viral infections, impacting host defense and viral replication.
• Understanding the dynamics of acetylation during infection is vital for deciphering virus-host interplay.

Purpose of the Study:

• To provide protocols for identifying, quantifying, and analyzing lysine acetylations during viral infections.
• To offer new perspectives on the mechanisms underlying viral pathogenesis through acetylation dynamics.
• To detail methods for assessing the cellular acetylome and site-specific acetylations.

Main Methods:

• Acetyl-lysine immunoaffinity purification coupled with quantitative mass spectrometry to analyze the cellular acetylome.
• Targeted mass spectrometry, specifically parallel reaction monitoring (PRM), for quantifying site-specific acetylations.
• Fluor de Lys assay for monitoring deacetylase activity.

Main Results:

• Established protocols for comprehensive analysis of lysine acetylation during viral infections.
• Demonstrated the utility of quantitative mass spectrometry and PRM for acetylation profiling.
• Provided a framework for functional characterization of acetylation events in the context of infection.

Conclusions:

• Detailed methodologies enable deeper investigation into the role of lysine acetylation in viral infections.
• These approaches facilitate the understanding of host-pathogen interactions at a molecular level.
• The study provides essential tools for advancing research in virology and epigenetics.