title: Studying the Lysine Acetylation of Malate Dehydrogenase.
abstract: Protein acetylation plays important roles in many biological processes. Malate dehydrogenase (MDH), a key enzyme in the tricarboxylic acid cycle, has been identified to be acetylated in bacteria by proteomic studies, but no further characterization has been reported. One challenge for studying protein acetylation is to get purely acetylated proteins at specific positions. Here, we applied the genetic code expansion strategy to site-specifically incorporate Nε-acetyllysine into MDH. The acetylation of lysine residues in MDH could enhance its enzyme activity. The Escherichia coli deacetylase CobB could deacetylate acetylated MDH, while the E. coli acetyltransferase YfiQ cannot acetylate MDH efficiently. Our results also demonstrated that acetyl-CoA or acetyl-phosphate could acetylate MDH chemically in vitro. Furthermore, the acetylation level of MDH was shown to be affected by carbon sources in the growth medium.
authors.affiliation: Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR 72701, USA; Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR 72701, USA.
authors.forename: Sumana
authors.initials: S
authors.lastname: Venkat
authors.affiliation: Department of Biological Sciences, University of Arkansas, Fayetteville, AR 72701, USA.
authors.forename: Caroline
authors.initials: C
authors.lastname: Gregory
authors.affiliation: Department of Biological Sciences, University of Arkansas, Fayetteville, AR 72701, USA.
authors.forename: Jourdan
authors.initials: J
authors.lastname: Sturges
authors.affiliation: Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR 72701, USA.
authors.forename: Qinglei
authors.initials: Q
authors.lastname: Gan
authors.affiliation: Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR 72701, USA; Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR 72701, USA. Electronic address: cf021@uark.edu.
authors.forename: Chenguang
authors.initials: C
authors.lastname: Fan
mesh_terms: D000105:Acetyl Coenzyme A / Q000378:metabolism; D000107:Acetylation; D000123:Acetyltransferases / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism; D008239:Lysine / Q000378:metabolism*; D008291:Malate Dehydrogenase / Q000378:metabolism*; D010755:Organophosphates / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D037761:Sirtuins / Q000378:metabolism
chemical_list: D003470:Culture Media; D029968:Escherichia coli Proteins; D010755:Organophosphates; C011632:acetyl phosphate; D000105:Acetyl Coenzyme A; D008291:Malate Dehydrogenase; D000123:Acetyltransferases; C561243:acetyl-CoA synthetase acetylase, E coli; D037761:Sirtuins; C483998:cobB protein, E Coli; D008239:Lysine
keywords: acetyltransferase; deacetylase; genetic code expansion; post-translational modification; protein acetylation