title: Nonenzymatic Protein Acetylation Detected by NAPPA Protein Arrays.
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 pKa 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.
authors.affiliation: Department of Biochemistry & Biophysics, Abramson Family Cancer Research Institute, Perelman School of Medicine at the University of Pennsylvania , Philadelphia, Pennsylvania, United States.|Program in Gene Expression and Regulation, The Wistar Institute , Philadelphia, Pennsylvania, United States.
authors.forename: Adam S
authors.initials: AS
authors.lastname: Olia
authors.affiliation: Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University , Tempe, Arizona, United States.
authors.forename: Kristi
authors.initials: K
authors.lastname: Barker
authors.affiliation: Department of Biochemistry & Biophysics, Abramson Family Cancer Research Institute, Perelman School of Medicine at the University of Pennsylvania , Philadelphia, Pennsylvania, United States.|Program in Gene Expression and Regulation, The Wistar Institute , Philadelphia, Pennsylvania, United States.
authors.forename: Cheryl E
authors.initials: CE
authors.lastname: McCullough
authors.affiliation: Program in Molecular and Cellular Oncogenesis, The Wistar Institute , Philadelphia, Pennsylvania, United States.
authors.forename: Hsin-Yao
authors.initials: HY
authors.lastname: Tang
authors.affiliation: Program in Molecular and Cellular Oncogenesis, The Wistar Institute , Philadelphia, Pennsylvania, United States.
authors.forename: David W
authors.initials: DW
authors.lastname: Speicher
authors.affiliation: Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University , Tempe, Arizona, United States.
authors.forename: Ji
authors.initials: J
authors.lastname: Qiu
authors.affiliation: Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University , Tempe, Arizona, United States.
authors.forename: Joshua
authors.initials: J
authors.lastname: LaBaer
authors.affiliation: Department of Biochemistry & Biophysics, Abramson Family Cancer Research Institute, Perelman School of Medicine at the University of Pennsylvania , Philadelphia, Pennsylvania, United States.|Program in Gene Expression and Regulation, The Wistar Institute , Philadelphia, Pennsylvania, United States.
authors.forename: Ronen
authors.initials: R
authors.lastname: Marmorstein
mesh_terms: D000107:Acetylation; D000595:Amino Acid Sequence; D006801:Humans; D008239:Lysine / Q000032:analysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D040081:Protein Array Analysis / Q000379:methods*; D011506:Proteins / Q000737:chemistry*
chemical_list: D011506:Proteins; D008239:Lysine