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Title: A Method to Determine Lysine Acetylation Stoichiometries

Lysine acetylation is a common protein posttranslational modification that regulates a variety of biological processes. A major bottleneck to fully understanding the functional aspects of lysine acetylation is the difficulty in measuring the proportion of lysine residues that are acetylated. Here we describe a mass spectrometry method using a combination of isotope labeling and detection of a diagnostic fragment ion to determine the stoichiometry of protein lysine acetylation. Using this technique, we determined the modification occupancy for ~750 acetylated peptides from mammalian cell lysates. Furthermore, the acetylation on N-terminal tail of histone H4 was cross-validated by treating cells with sodium butyrate, a potent deacetylase inhibitor, and comparing changes in stoichiometry levels measured by our method with immunoblotting measurements. Of note we observe that acetylation stoichiometry is high in nuclear proteins, but very low in mitochondrial and cytosolic proteins. In summary, our method opens new opportunities to study in detail the relationship of lysine acetylation levels of proteins with their biological functions.
Authors:
 [1] ;  [2] ;  [3] ;  [3] ;  [3] ;  [3] ;  [2] ;  [4] ;  [3] ;  [3] ;  [2] ;  [3] ;  [3] ;  [3] ;  [3]
  1. Biological Science Division and Environmental, Pacific Northwest National Laboratory, Richland, WA 99352, USA, Bindley Bioscience Center, Discovery Park, Purdue University, West Lafayette, IN 47907, USA
  2. Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99352, USA
  3. Biological Science Division and Environmental, Pacific Northwest National Laboratory, Richland, WA 99352, USA
  4. Biological Science Division and Environmental, Pacific Northwest National Laboratory, Richland, WA 99352, USA, Institute for Basic Science, Seoul National University, Seoul 151-747, Republic of Korea
Publication Date:
OSTI Identifier:
1198079
Grant/Contract Number:
AC05-76-RLO1830
Type:
Published Article
Journal Name:
International Journal of Proteomics
Additional Journal Information:
Journal Volume: 2014; Related Information: CHORUS Timestamp: 2016-07-29 02:35:38; Journal ID: ISSN 2090-2166
Publisher:
Hindawi Publishing Corporation
Sponsoring Org:
USDOE
Country of Publication:
Country unknown/Code not available
Language:
English