Metabolomics analysis identifies d-alanine-d-alanine ligase as the primary lethal target of d-cycloserine in mycobacteria

Steven Halouska, Robert J. Fenton, Denise K. Zinniel, Darrell D. Marshall, Raul G Barletta, Robert Powers

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

d-Cycloserine is an effective second line antibiotic used as a last resort to treat multi (MDR)- and extensively (XDR) drug resistant strains of Mycobacterium tuberculosis. d-Cycloserine interferes with the formation of peptidoglycan biosynthesis by competitive inhibition of alanine racemase (Alr) and d-alanine-d-alanine ligase (Ddl). Although the two enzymes are known to be inhibited, the in vivo lethal target is still unknown. Our NMR metabolomics work has revealed that Ddl is the primary target of DCS, as cell growth is inhibited when the production of d-alanyl-d-alanine is halted. It is shown that inhibition of Alr may contribute indirectly by lowering the levels of d-alanine, thus allowing DCS to outcompete d-alanine for Ddl binding. The NMR data also supports the possibility of a transamination reaction to produce d-alanine from pyruvate and glutamate, thereby bypassing Alr inhibition. Furthermore, the inhibition of peptidoglycan synthesis results in a cascading effect on cellular metabolism as there is a shift toward the catabolic routes to compensate for accumulation of peptidoglycan precursors.

Original languageEnglish (US)
Pages (from-to)1065-1076
Number of pages12
JournalJournal of proteome research
Volume13
Issue number2
DOIs
StatePublished - Feb 7 2014

Fingerprint

Cycloserine
Metabolomics
Mycobacterium
Ligases
Alanine
Alanine Racemase
Peptidoglycan
Nuclear magnetic resonance
Biosynthesis
Cell growth
Pyruvic Acid
Mycobacterium tuberculosis
Metabolism
Glutamic Acid

Keywords

  • NMR metabolomics
  • d -cycloserine
  • mycobacteria
  • tuberculosis

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Metabolomics analysis identifies d-alanine-d-alanine ligase as the primary lethal target of d-cycloserine in mycobacteria. / Halouska, Steven; Fenton, Robert J.; Zinniel, Denise K.; Marshall, Darrell D.; Barletta, Raul G; Powers, Robert.

In: Journal of proteome research, Vol. 13, No. 2, 07.02.2014, p. 1065-1076.

Research output: Contribution to journalArticle

Halouska, Steven ; Fenton, Robert J. ; Zinniel, Denise K. ; Marshall, Darrell D. ; Barletta, Raul G ; Powers, Robert. / Metabolomics analysis identifies d-alanine-d-alanine ligase as the primary lethal target of d-cycloserine in mycobacteria. In: Journal of proteome research. 2014 ; Vol. 13, No. 2. pp. 1065-1076.
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