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Divergence of multimodular polyketide synthases revealed by a didomain structure

Nature Chemical Biology volume 8, pages 615–621 (2012)Cite this article

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Abstract

The enoylreductase (ER) is the final common enzyme from modular polyketide synthases (PKSs) to be structurally characterized. The 3.0 Å–resolution structure of the didomain comprising the ketoreductase (KR) and ER from the second module of the spinosyn PKS reveals that ER shares an ∼600-Å2 interface with KR distinct from that of the related mammalian fatty acid synthase (FAS). In contrast to the ER domains of the mammalian FAS, the ER domains of the second module of the spinosyn PKS do not make contact across the two-fold axis of the synthase. This monomeric organization may have been necessary in the evolution of multimodular PKSs to enable acyl carrier proteins to access each of their cognate enzymes. The isolated ER domain showed activity toward a substrate analog, enabling us to determine the contributions of its active site residues.

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Figure 1: A complete module from the spinosyn PKS.
Figure 2: KR and ER architecture.
Figure 3: The structure of SpnER2.
Figure 4: Functional analysis of SpnER2.
Figure 5: Synthase schematic.

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Acknowledgements

We thank A.F. Monzingo for helping with in-house diffraction experiments and C.D. Fage for helping optimize crystals. Synchrotron data were obtained at the ALS Beamlines 8.2.1 and 5.0.2. Financial support was provided by Welch Foundation Grant F-1712 (A.T.K.-C.) as well as the Sealy and Smith Foundation grant to the Sealy Center for Structural Biology and Molecular Biophysics (M.A.W.). The development of the UltraScan software is supported by the US National Institutes of Health through grant RR022200 (B.D.). Supercomputer time allocations were provided through the US National Science Foundation grant TG-MCB070038 (B.D.). We acknowledge the support of the San Antonio Cancer Institute grant P30 CA054174 for the Center for Analytical Ultracentrifugation of Macromolecular Assemblies at the University of Texas Health Science Center at San Antonio.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, The University of Texas at Austin, Austin, Texas, USA

    Jianting Zheng, Darren C Gay & Adrian T Keatinge-Clay

  2. Department of Biochemistry, The University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA

    Borries Demeler

  3. Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch at Galveston, Galveston, Texas, USA

    Mark A White

  4. Department of Biochemistry and Molecular Biology, University of Texas Medical Branch at Galveston, Galveston, Texas, USA

    Mark A White

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  1. Jianting Zheng
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Contributions

J.Z. performed all of the studies except for the functional assays (conducted by D.C.G.), analytical ultracentrifugation (conducted by B.D.) and SAXS (conducted by M.A.W.). All authors contributed to writing the manuscript and creating the figures.

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Correspondence to Adrian T Keatinge-Clay.

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Zheng, J., Gay, D., Demeler, B. et al. Divergence of multimodular polyketide synthases revealed by a didomain structure. Nat Chem Biol 8, 615–621 (2012). https://doi.org/10.1038/nchembio.964

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