Characterization of Lipoyl Synthase from Mycobacterium tuberculosis

Nicholas D. Lanz, Kyung Hoon Lee, Abigail K. Horstmann, Maria Eirini Pandelia, Robert M. Cicchillo, Carsten Krebs, Squire J. Booker

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The prevalence of multiple and extensively drug-resistant strains of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, is on the rise, necessitating the identification of new targets to combat an organism that has infected one-third of the world's population, according to the World Health Organization. The biosynthesis of the lipoyl cofactor is one possible target, given its critical importance in cellular metabolism and the apparent lack of functional salvage pathways in Mtb that are found in humans and many other organisms. The lipoyl cofactor is synthesized de novo in two committed steps, involving the LipB-catalyzed transfer of an octanoyl chain derived from fatty acid biosynthesis to a lipoyl carrier protein and the LipA-catalyzed insertion of sulfur atoms at C6 and C8 of the octanoyl chain. A number of in vitro studies of lipoyl synthases from Escherichia coli, Sulfolobus solfataricus, and Thermosynechococcus elongatus have been conducted, but the enzyme from Mtb has not been characterized. Herein, we show that LipA from Mtb contains two [4Fe-4S] clusters and converts an octanoyl peptide substrate to the corresponding lipoyl peptide product via the same C6-monothiolated intermediate as that observed in the E. coli LipA reaction. In addition, we show that LipA from Mtb forms a complex with the H protein of the glycine cleavage system and that the strength of association is dependent on the presence of S-adenosyl-l-methionine. We also show that LipA from Mtb can complement a lipA mutant of E. coli, demonstrating the commonalities of the two enzymes. Lastly, we show that the substrate for LipA, which normally acts on a post-translationally modified protein, can be reduced to carboxybenzyl-octanoyllysine.

Original languageEnglish (US)
Pages (from-to)1372-1383
Number of pages12
JournalBiochemistry
Volume55
Issue number9
DOIs
StatePublished - Mar 8 2016

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Mycobacterium tuberculosis
Escherichia coli
Biosynthesis
Glycine Decarboxylase Complex H-Protein
Salvaging
Peptides
Substrates
Enzymes
Sulfur
Metabolism
Methionine
Carrier Proteins
Fatty Acids
Health
Association reactions
Sulfolobus solfataricus
Atoms
Pharmaceutical Preparations
Tuberculosis
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Lanz, N. D., Lee, K. H., Horstmann, A. K., Pandelia, M. E., Cicchillo, R. M., Krebs, C., & Booker, S. J. (2016). Characterization of Lipoyl Synthase from Mycobacterium tuberculosis. Biochemistry, 55(9), 1372-1383. https://doi.org/10.1021/acs.biochem.5b01216
Lanz, Nicholas D. ; Lee, Kyung Hoon ; Horstmann, Abigail K. ; Pandelia, Maria Eirini ; Cicchillo, Robert M. ; Krebs, Carsten ; Booker, Squire J. / Characterization of Lipoyl Synthase from Mycobacterium tuberculosis. In: Biochemistry. 2016 ; Vol. 55, No. 9. pp. 1372-1383.
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Lanz, ND, Lee, KH, Horstmann, AK, Pandelia, ME, Cicchillo, RM, Krebs, C & Booker, SJ 2016, 'Characterization of Lipoyl Synthase from Mycobacterium tuberculosis', Biochemistry, vol. 55, no. 9, pp. 1372-1383. https://doi.org/10.1021/acs.biochem.5b01216

Characterization of Lipoyl Synthase from Mycobacterium tuberculosis. / Lanz, Nicholas D.; Lee, Kyung Hoon; Horstmann, Abigail K.; Pandelia, Maria Eirini; Cicchillo, Robert M.; Krebs, Carsten; Booker, Squire J.

In: Biochemistry, Vol. 55, No. 9, 08.03.2016, p. 1372-1383.

Research output: Contribution to journalArticle

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AU - Lanz, Nicholas D.

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Lanz ND, Lee KH, Horstmann AK, Pandelia ME, Cicchillo RM, Krebs C et al. Characterization of Lipoyl Synthase from Mycobacterium tuberculosis. Biochemistry. 2016 Mar 8;55(9):1372-1383. https://doi.org/10.1021/acs.biochem.5b01216