Site-Specific Lysine Acetylation Stoichiometry Across Subcellular Compartments.

Area of Science:

• Proteomics
• Molecular Biology
• Biochemistry

Background:

• Posttranslational protein modifications regulate critical cellular processes.
• Protein acetylation is a widespread, dynamic modification with incompletely understood regulation and effects.
• Accurate assessment of acetylation stoichiometry is needed.

Purpose of the Study:

• To determine protein acetylation stoichiometry.
• To expand the number of quantified acetylation sites.
• To gain insights into compartment-specific regulation and functions of protein acetylation.

Main Methods:

• Utilized data-independent acquisition mass spectrometry for quantification.
• Employed subcellular fractionation and offline prefractionation to enhance spectral library depth.
• Interrogated subcellular compartments for deep analysis.

Main Results:

• Expanded the number of quantified protein acetylation sites.
• Improved the depth of data-independent acquisition by enhancing the spectral library.
• Enabled deep interrogation of subcellular compartments.

Conclusions:

• The described method allows for a more comprehensive determination of acetylation stoichiometry.
• Enhanced spectral libraries through fractionation improve proteomic data analysis.
• This approach provides crucial insights into the compartment-specific roles of protein acetylation.