title: Lysine acetylation regulates the activity of Escherichia coli pyridoxine 5'-phosphate oxidase.
abstract: Nɛ-lysine acetylation is one of the most abundant post-translational modifications in eukaryote and prokaryote. Protein acetylome of Escherichia coli has been screened using mass spectrometry (MS) technology, and many acetylated proteins have been identified, including the pyridoxine 5'-phosphate oxidase (EcPNPOx), but the biological roles played by lysine acetylation in EcPNPOx still remain unknown. In this study, EcPNPOx was firstly overexpressed and purified, and two acetylated lysine residues were identified by the subsequent liquid chromatography-tandem mass spectrometry analysis. Site-directed mutagenesis analysis demonstrated that acetylated lysine residues play important roles in the enzymatic activity and enzymatic properties of the protein. EcPNPOx could be non-enzymatically acetylated by acetyl-phosphate and deacetylated by CobB in vitro. Furthermore, enzymatic activities of acetylated and deacetylated EcPNPOx were compared in vitro, and results showed that acetylation led to a decrease of its enzymatic activity, which could be rescued by CobB deacetylation. Taken together, our data suggest that CobB modulates the enzymatic activity of EcPNPOx in vitro.
authors.affiliation: Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Gu
authors.affiliation: Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.|Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.|Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Hongsen
authors.initials: H
authors.lastname: Guo
authors.affiliation: Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.|University of Chinese Academy of Sciences, Beijing 100039, China.
authors.forename: Manluan
authors.initials: M
authors.lastname: Sun
authors.affiliation: Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430071, China.
authors.forename: Mingkun
authors.initials: M
authors.lastname: Yang
authors.affiliation: Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
authors.forename: Xude
authors.initials: X
authors.lastname: Wang
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Xian'en
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
authors.forename: Jiaoyu
authors.initials: J
authors.lastname: Deng
mesh_terms: D000107:Acetylation; D000595:Amino Acid Sequence; D015153:Blotting, Western; D002853:Chromatography, Liquid; D057075:Enzyme Assays / Q000379:methods; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008239:Lysine / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D011499:Protein Processing, Post-Translational; D011734:Pyridoxaminephosphate Oxidase / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D037761:Sirtuins / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D013696:Temperature
chemical_list: D029968:Escherichia coli Proteins; D011734:Pyridoxaminephosphate Oxidase; D037761:Sirtuins; C483998:cobB protein, E Coli; D008239:Lysine
keywords: CobB; deacetylation; lysine acetylation; pyridoxine 5′-phosphate oxidase