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Characterizing Lysine Acetylation of Isocitrate Dehydrogenase in Escherichia coli.
J Mol Biol. 2018 06 22; 430(13):1901-1911.JM

Abstract

The Escherichia coli isocitrate dehydrogenase (ICDH) is one of the tricarboxylic acid cycle enzymes, playing key roles in energy production and carbon flux regulation. E. coli ICDH was the first bacterial enzyme shown to be regulated by reversible phosphorylation. However, the effect of lysine acetylation on E. coli ICDH, which has no sequence similarity with its counterparts in eukaryotes, is still unclear. Based on previous studies of E. coli acetylome and ICDH crystal structures, eight lysine residues were selected for mutational and kinetic analyses. They were replaced with acetyllysine by the genetic code expansion strategy or substituted with glutamine as a classic approach. Although acetylation decreased the overall ICDH activity, its effects were different site by site. Deacetylation tests demonstrated that the CobB deacetylase could deacetylate ICDH both in vivo and in vitro, but CobB was only specific for lysine residues at the protein surface. On the other hand, ICDH could be acetylated by acetyl-phosphate chemically in vitro. And in vivo acetylation tests indicated that the acetylation level of ICDH was correlated with the amounts of intracellular acetyl-phosphate. This study nicely complements previous proteomic studies to provide direct biochemical evidence for ICDH acetylation.

Authors+Show Affiliations

Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR 72701, United States; Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR 72701, United States.Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR 72701, United States; Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR 72701, United States.Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR 72701, United States.Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR 72701, United States.Department of Biological Sciences, University of Arkansas, Fayetteville, AR 72701, United States.Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR 72701, United States.Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR 72701, United States; Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR 72701, United States. Electronic address: cf021@uark.edu.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

29733852

Citation

Venkat, Sumana, et al. "Characterizing Lysine Acetylation of Isocitrate Dehydrogenase in Escherichia Coli." Journal of Molecular Biology, vol. 430, no. 13, 2018, pp. 1901-1911.
Venkat S, Chen H, Stahman A, et al. Characterizing Lysine Acetylation of Isocitrate Dehydrogenase in Escherichia coli. J Mol Biol. 2018;430(13):1901-1911.
Venkat, S., Chen, H., Stahman, A., Hudson, D., McGuire, P., Gan, Q., & Fan, C. (2018). Characterizing Lysine Acetylation of Isocitrate Dehydrogenase in Escherichia coli. Journal of Molecular Biology, 430(13), 1901-1911. https://doi.org/10.1016/j.jmb.2018.04.031
Venkat S, et al. Characterizing Lysine Acetylation of Isocitrate Dehydrogenase in Escherichia Coli. J Mol Biol. 2018 06 22;430(13):1901-1911. PubMed PMID: 29733852.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterizing Lysine Acetylation of Isocitrate Dehydrogenase in Escherichia coli. AU - Venkat,Sumana, AU - Chen,Hao, AU - Stahman,Alleigh, AU - Hudson,Denver, AU - McGuire,Paige, AU - Gan,Qinglei, AU - Fan,Chenguang, Y1 - 2018/05/04/ PY - 2018/03/18/received PY - 2018/04/18/revised PY - 2018/04/24/accepted PY - 2018/5/8/pubmed PY - 2019/5/29/medline PY - 2018/5/8/entrez KW - carbon flux KW - genetic code expansion KW - glyoxylate bypass KW - post-translational modification KW - tricarboxylic acid cycle SP - 1901 EP - 1911 JF - Journal of molecular biology JO - J Mol Biol VL - 430 IS - 13 N2 - The Escherichia coli isocitrate dehydrogenase (ICDH) is one of the tricarboxylic acid cycle enzymes, playing key roles in energy production and carbon flux regulation. E. coli ICDH was the first bacterial enzyme shown to be regulated by reversible phosphorylation. However, the effect of lysine acetylation on E. coli ICDH, which has no sequence similarity with its counterparts in eukaryotes, is still unclear. Based on previous studies of E. coli acetylome and ICDH crystal structures, eight lysine residues were selected for mutational and kinetic analyses. They were replaced with acetyllysine by the genetic code expansion strategy or substituted with glutamine as a classic approach. Although acetylation decreased the overall ICDH activity, its effects were different site by site. Deacetylation tests demonstrated that the CobB deacetylase could deacetylate ICDH both in vivo and in vitro, but CobB was only specific for lysine residues at the protein surface. On the other hand, ICDH could be acetylated by acetyl-phosphate chemically in vitro. And in vivo acetylation tests indicated that the acetylation level of ICDH was correlated with the amounts of intracellular acetyl-phosphate. This study nicely complements previous proteomic studies to provide direct biochemical evidence for ICDH acetylation. SN - 1089-8638 UR - https://www.unboundmedicine.com/medline/citation/29733852/Characterizing_Lysine_Acetylation_of_Isocitrate_Dehydrogenase_in_Escherichia_coli_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(18)30342-5 DB - PRIME DP - Unbound Medicine ER -