Abstract
The emerging view of Nε-lysine acetylation in eukaryotes is of a relatively abundant post-translational modification (PTM) that has a major impact on the function, structure, stability and/or location of thousands of proteins involved in diverse cellular processes. This PTM is typically considered to arise by the donation of the acetyl group from acetyl-coenzyme A (acCoA) to the ε-amino group of a lysine residue that is reversibly catalyzed by lysine acetyltransferases and deacetylases. Here, we provide genetic, mass spectrometric, biochemical and structural evidence that N ε-lysine acetylation is an equally abundant and important PTM in bacteria. Applying a recently developed, label-free and global mass spectrometric approach to an isogenic set of mutants, we detected acetylation of thousands of lysine residues on hundreds of Escherichia coli proteins that participate in diverse and often essential cellular processes, including translation, transcription and central metabolism. Many of these acetylations were regulated in an acetyl phosphate (acP)-dependent manner, providing compelling evidence for a recently reported mechanism of bacterial N ε-lysine acetylation. These mass spectrometric data, coupled with observations made by crystallography, biochemistry, and additional mass spectrometry showed that this acP-dependent acetylation is both non-enzymatic and specific, with specificity determined by the accessibility, reactivity and three-dimensional microenvironment of the target lysine. Crystallographic evidence shows acP can bind to proteins in active sites and cofactor binding sites, but also potentially anywhere molecules with a phosphate moiety could bind. Finally, we provide evidence that acP-dependent acetylation can impact the function of critical enzymes, including glyceraldehyde-3-phosphate dehydrogenase, triosephosphate isomerase, and RNA polymerase.
Original language | English (US) |
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Article number | e94816 |
Journal | PloS one |
Volume | 9 |
Issue number | 4 |
DOIs | |
State | Published - Apr 22 2014 |
ASJC Scopus subject areas
- General
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Dive into the research topics of 'Structural, kinetic and proteomic characterization of acetyl phosphate-dependent bacterial protein acetylation'. Together they form a unique fingerprint.Datasets
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1.43 Angstrom Resolution Crystal Structure of Triosephosphate Isomerase (tpiA) from Escherichia coli in Complex with Acetyl Phosphate.
Kuhn, M. L. (Contributor), Zemaitaitis, B. (Contributor), Hu, L. I. (Contributor), Sahu, A. (Contributor), Sorensen, D. (Contributor), Minasov, G. (Contributor), Lima, B. P. (Contributor), Scholle, M. (Contributor), Mrksich, M. (Contributor), Anderson, W. F. (Contributor), Gibson, B. W. (Contributor), Schilling, B. (Contributor) & Wolfe, A. J. (Contributor), Protein Data Bank (PDB), Apr 16 2014
DOI: 10.2210/pdb4MVA/pdb, https://www.wwpdb.org/pdb?id=pdb_00004mva
Dataset
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Revised Crystal Structure of apo-form of Triosephosphate Isomerase (tpiA) from Escherichia coli at 1.8 Angstrom Resolution.
Kuhn, M. L. (Contributor), Zemaitaitis, B. (Contributor), Hu, L. I. (Contributor), Sahu, A. (Contributor), Sorensen, D. (Contributor), Minasov, G. (Contributor), Lima, B. P. (Contributor), Scholle, M. (Contributor), Mrksich, M. (Contributor), Anderson, W. F. (Contributor), Gibson, B. W. (Contributor), Schilling, B. (Contributor) & Wolfe, A. J. (Contributor), Protein Data Bank (PDB), May 1 2013
DOI: 10.2210/pdb4K6A/pdb, https://www.wwpdb.org/pdb?id=pdb_00004k6a
Dataset
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2.85 Angstrom Resolution Crystal Structure of Glyceraldehyde 3-phosphate Dehydrogenase A (gapA) from Escherichia coli Modified by Acetyl Phosphate.
Kuhn, M. L. (Contributor), Zemaitaitis, B. (Contributor), Hu, L. I. (Contributor), Sahu, A. (Contributor), Sorensen, D. (Contributor), Minasov, G. (Contributor), Lima, B. P. (Contributor), Scholle, M. (Contributor), Mrksich, M. (Contributor), Anderson, W. F. (Contributor), Gibson, B. W. (Contributor), Schilling, B. (Contributor) & Wolfe, A. J. (Contributor), Protein Data Bank (PDB), Apr 23 2014
DOI: 10.2210/pdb4MVJ/pdb, https://www.wwpdb.org/pdb?id=pdb_00004mvj
Dataset