Crystal structure of Mycobacterium tuberculosis polyketide synthase 11 (PKS11) reveals intermediates in the synthesis of methyl-branched alkylpyrones

Kuppan Gokulan, Seán E. O'Leary, William Russell, David H. Russell, Mallikarjun Lalgondar, Tadhg P. Begley, Thomas R. Ioerger, James C. Sacchettini

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

PKS11 is one of three type III polyketide synthases (PKSs) identified in Mycobacterium tuberculosis. Although many PKSs in M. tuberculosis have been implicated in producing complex cell wall glycolipids, the biological function of PKS11 is unknown. PKS11 has previously been proposed to synthesize alkylpyrones from fatty acid substrates. We solved the crystal structure of M. tuberculosis PKS11 and found the overall fold to be similar to other type III PKSs. PKS11 has a deep hydrophobic tunnel proximal to the active site Cys-138 to accommodate substrates. We observed electron density in this tunnel from a co-purified molecule that was identified by mass spectrometry to be palmitate. Co-crystallization with malonyl-CoA (MCoA) or methylmalonyl-CoA (MMCoA) led to partial turnover of the substrate, resulting in trapped intermediates. Reconstitution of the reaction in solution confirmed that both co-factors are required for optimal activity, and kinetic analysis shows that MMCoA is incorporated first, then MCoA, followed by lactonization to produce methyl-branched alkylpyrones.

Original languageEnglish (US)
Pages (from-to)16484-16494
Number of pages11
JournalJournal of Biological Chemistry
Volume288
Issue number23
DOIs
StatePublished - Jun 7 2013
Externally publishedYes

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Polyketide Synthases
Mycobacterium tuberculosis
Crystal structure
Malonyl Coenzyme A
Tunnels
Substrates
Palmitates
Glycolipids
Crystallization
Cell Wall
Carrier concentration
Mass spectrometry
Mass Spectrometry
Catalytic Domain
Fatty Acids
Cells
Electrons
Molecules
Kinetics

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Crystal structure of Mycobacterium tuberculosis polyketide synthase 11 (PKS11) reveals intermediates in the synthesis of methyl-branched alkylpyrones. / Gokulan, Kuppan; O'Leary, Seán E.; Russell, William; Russell, David H.; Lalgondar, Mallikarjun; Begley, Tadhg P.; Ioerger, Thomas R.; Sacchettini, James C.

In: Journal of Biological Chemistry, Vol. 288, No. 23, 07.06.2013, p. 16484-16494.

Research output: Contribution to journalArticle

Gokulan, Kuppan ; O'Leary, Seán E. ; Russell, William ; Russell, David H. ; Lalgondar, Mallikarjun ; Begley, Tadhg P. ; Ioerger, Thomas R. ; Sacchettini, James C. / Crystal structure of Mycobacterium tuberculosis polyketide synthase 11 (PKS11) reveals intermediates in the synthesis of methyl-branched alkylpyrones. In: Journal of Biological Chemistry. 2013 ; Vol. 288, No. 23. pp. 16484-16494.
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