PMID: 2121129Oct 1, 1990Paper

Isolation and characterization of lipoylated and unlipoylated domains of the E2p subunit of the pyruvate dehydrogenase complex of Escherichia coli

The Biochemical Journal
S T Ali, J R Guest

Abstract

The dihydrolipoamide acetyltransferase subunit (E2p) of the pyruvate dehydrogenase complex of Escherichia coli has three highly conserved and tandemly repeated lipoyl domains, each containing approx. 80 amino acid residues. These domains are covalently modified with lipoyl groups bound in amide linkage to the N6-amino groups of specific lysine residues, and the cofactors perform essential roles in the formation and transfer of acetyl groups by the dehydrogenase (E1p) and acetyltransferase (E2p) subunits. A subgene encoding a hybrid lipoyl domain was previously shown to generate two products when overexpressed, whereas a mutant subgene, in which the lipoyl-lysine codon is replaced by a glutamine codon, expresses only one product. A method has been devised for purifying the three types of independently folded domain from crude extracts of E. coli, based on their pH-(and heat-)stabilities. The domains were characterized by: amino acid and N-terminal sequence analysis, lipoic acid content, acetylation by E1p, tryptic peptide analysis and immunochemical activity. This has shown that the two forms of domain expressed from the parental subgene are lipoylated (L203) and unlipoylated (U203) derivatives of the hybrid lipoyl domain, where...Continue Reading

Citations

Jan 1, 2004·Chemistry & Biology·Xin ZhaoJohn E Cronan
Jan 10, 2017·PloS One·Yirong SunHaihong Wang
Jan 18, 2018·Proceedings of the National Academy of Sciences of the United States of America·Xinyun CaoJohn E Cronan
Dec 16, 1991·FEBS Letters·D E BrookfieldJ R Guest
Aug 1, 1995·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·P S LeungM E Gershwin
Apr 15, 2016·Microbiology and Molecular Biology Reviews : MMBR·John E Cronan
Dec 7, 2007·Nature Biotechnology·Marta Fernández-SuárezAlice Y Ting
Jan 7, 2011·The Journal of Biological Chemistry·Bachar H Hassan, John E Cronan
Oct 6, 2021·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Yao YangM Eric Gershwin

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