A Method to Determine Lysine Acetylation Stoichiometries

Nakayasu, Ernesto S.; Wu, Si; Sydor, Michael A.; Shukla, Anil K.; Weitz, Karl K.; Moore, Ronald J.; Hixson, Kim K.; Kim, Jong-Seo; Petyuk, Vladislav A.; Monroe, Matthew E.; Pasa-Tolic, Ljiljiana; Qian, Wei-Jun; Smith, Richard D.; Adkins, Joshua N.; Ansong, Charles

doi:10.1155/2014/730725

Title: A Method to Determine Lysine Acetylation Stoichiometries

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Abstract

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:

Nakayasu, Ernesto S. ^[1]; Wu, Si ^[2]; Sydor, Michael A. ^[3]; Shukla, Anil K. ^[3]; Weitz, Karl K. ^[3]; Moore, Ronald J. ^[3]; Hixson, Kim K. ^[2]; Kim, Jong-Seo ^[4]; Petyuk, Vladislav A. ^[3]; Monroe, Matthew E. ^[3]; Pasa-Tolic, Ljiljiana ^[2]; Qian, Wei-Jun ^[3]; Smith, Richard D. ^[3]; Adkins, Joshua N. ^[3]; Ansong, Charles ^[3]

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
Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99352, USA
Biological Science Division and Environmental, Pacific Northwest National Laboratory, Richland, WA 99352, USA
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:: Wed Jan 01 00:00:00 EST 2014

Sponsoring Org.:: USDOE

OSTI Identifier:: 1198079

Grant/Contract Number:: AC05-76-RLO1830

Resource Type:: Published Article

Journal Name:: International Journal of Proteomics

Additional Journal Information:: Journal Name: International Journal of Proteomics Journal Volume: 2014; Journal ID: ISSN 2090-2166

Publisher:: Hindawi Publishing Corporation

Country of Publication:: Country unknown/Code not available

Language:: English

Citation Formats


                    Nakayasu, Ernesto S., Wu, Si, Sydor, Michael A., Shukla, Anil K., Weitz, Karl K., Moore, Ronald J., Hixson, Kim K., Kim, Jong-Seo, Petyuk, Vladislav A., Monroe, Matthew E., Pasa-Tolic, Ljiljiana, Qian, Wei-Jun, Smith, Richard D., Adkins, Joshua N., and Ansong, Charles. A Method to Determine Lysine Acetylation Stoichiometries.  Country unknown/Code not available: N. p., 2014. 
Web.  doi:10.1155/2014/730725.

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                    Nakayasu, Ernesto S., Wu, Si, Sydor, Michael A., Shukla, Anil K., Weitz, Karl K., Moore, Ronald J., Hixson, Kim K., Kim, Jong-Seo, Petyuk, Vladislav A., Monroe, Matthew E., Pasa-Tolic, Ljiljiana, Qian, Wei-Jun, Smith, Richard D., Adkins, Joshua N., & Ansong, Charles. A Method to Determine Lysine Acetylation Stoichiometries.  Country unknown/Code not available.  https://doi.org/10.1155/2014/730725

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                    Nakayasu, Ernesto S., Wu, Si, Sydor, Michael A., Shukla, Anil K., Weitz, Karl K., Moore, Ronald J., Hixson, Kim K., Kim, Jong-Seo, Petyuk, Vladislav A., Monroe, Matthew E., Pasa-Tolic, Ljiljiana, Qian, Wei-Jun, Smith, Richard D., Adkins, Joshua N., and Ansong, Charles. Wed .  
"A Method to Determine Lysine Acetylation Stoichiometries".  Country unknown/Code not available.  https://doi.org/10.1155/2014/730725.

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@article{osti_1198079,

  title        = {A Method to Determine Lysine Acetylation Stoichiometries},

  author       = {Nakayasu, Ernesto S. and Wu, Si and Sydor, Michael A. and Shukla, Anil K. and Weitz, Karl K. and Moore, Ronald J. and Hixson, Kim K. and Kim, Jong-Seo and Petyuk, Vladislav A. and Monroe, Matthew E. and Pasa-Tolic, Ljiljiana and Qian, Wei-Jun and Smith, Richard D. and Adkins, Joshua N. and Ansong, Charles},

  abstractNote = {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.},

  doi          = {10.1155/2014/730725},

  journal      = {International Journal of Proteomics},

  number       = ,

  volume       = 2014,

  place        = {Country unknown/Code not available},

  year         = {Wed Jan 01 00:00:00 EST 2014},

  month        = {Wed Jan 01 00:00:00 EST 2014}

}

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