Nonenzymatic Protein Acetylation Detected by NAPPA Protein Arrays

Adam S. Olia, Kristi Barker, Cheryl E. McCullough, Hsin Yao Tang, David W. Speicher, Ji Qiu, Joshua LaBaer, Ronen Marmorstein

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Acetylation is a post-translational modification that occurs on thousands of proteins located in many cellular organelles. This process mediates many protein functions and modulates diverse biological processes. In mammalian cells, where acetyl-CoA is the primary acetyl donor, acetylation in the mitochondria is thought to occur by chemical means due to the relatively high concentration of acetyl-CoA located in this organelle. In contrast, acetylation outside of the mitochondria is thought to be mediated predominantly by acetyltransferase enzymes. Here, we address the possibility that nonenzymatic chemical acetylation outside of the mitochondria may be more common than previously appreciated. We employed the Nucleic Acid Programmable Protein Array platform to perform an unbiased screen for human proteins that undergo chemical acetylation, which resulted in the identification of a multitude of proteins with diverse functions and cellular localization. Mass spectrometry analysis revealed that basic residues typically precede the acetylated lysine in the -7 to -3 position, and we show by mutagenesis that these basic residues contribute to chemical acetylation capacity. We propose that these basic residues lower the pK<inf>a</inf> of the substrate lysine for efficient chemical acetylation. Many of the identified proteins reside outside of the mitochondria and have been previously demonstrated to be acetylated in vivo. As such, our studies demonstrate that chemical acetylation occurs more broadly throughout the eukaryotic cell than previously appreciated and suggests that this post-translational protein modification may have more diverse roles in protein function and pathway regulation.

Original languageEnglish (US)
Pages (from-to)2034-2047
Number of pages14
JournalACS Chemical Biology
Volume10
Issue number9
DOIs
StatePublished - Sep 18 2015

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Acetylation
Protein Array Analysis
Mitochondria
Proteins
Acetyl Coenzyme A
Post Translational Protein Processing
Organelles
Lysine
Biological Phenomena
Mutagenesis
Acetyltransferases
Eukaryotic Cells
Nucleic Acids
Mass spectrometry
Mass Spectrometry
Cells
Substrates
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

Olia, A. S., Barker, K., McCullough, C. E., Tang, H. Y., Speicher, D. W., Qiu, J., ... Marmorstein, R. (2015). Nonenzymatic Protein Acetylation Detected by NAPPA Protein Arrays. ACS Chemical Biology, 10(9), 2034-2047. https://doi.org/10.1021/acschembio.5b00342

Nonenzymatic Protein Acetylation Detected by NAPPA Protein Arrays. / Olia, Adam S.; Barker, Kristi; McCullough, Cheryl E.; Tang, Hsin Yao; Speicher, David W.; Qiu, Ji; LaBaer, Joshua; Marmorstein, Ronen.

In: ACS Chemical Biology, Vol. 10, No. 9, 18.09.2015, p. 2034-2047.

Research output: Contribution to journalArticle

Olia, AS, Barker, K, McCullough, CE, Tang, HY, Speicher, DW, Qiu, J, LaBaer, J & Marmorstein, R 2015, 'Nonenzymatic Protein Acetylation Detected by NAPPA Protein Arrays', ACS Chemical Biology, vol. 10, no. 9, pp. 2034-2047. https://doi.org/10.1021/acschembio.5b00342
Olia AS, Barker K, McCullough CE, Tang HY, Speicher DW, Qiu J et al. Nonenzymatic Protein Acetylation Detected by NAPPA Protein Arrays. ACS Chemical Biology. 2015 Sep 18;10(9):2034-2047. https://doi.org/10.1021/acschembio.5b00342
Olia, Adam S. ; Barker, Kristi ; McCullough, Cheryl E. ; Tang, Hsin Yao ; Speicher, David W. ; Qiu, Ji ; LaBaer, Joshua ; Marmorstein, Ronen. / Nonenzymatic Protein Acetylation Detected by NAPPA Protein Arrays. In: ACS Chemical Biology. 2015 ; Vol. 10, No. 9. pp. 2034-2047.
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