Abstract
Many important natural products are produced by multidomain non-ribosomal peptide synthetases (NRPSs). During synthesis, intermediates are covalently bound to integrated carrier domains and transported to neighbouring catalytic domains in an assembly line fashion. Understanding the structural basis for catalysis with non-ribosomal peptide synthetases will facilitate bioengineering to create novel products. Here we describe the structures of two different holo-non-ribosomal peptide synthetase modules, each revealing a distinct step in the catalytic cycle. One structure depicts the carrier domain cofactor bound to the peptide bond-forming condensation domain, whereas a second structure captures the installation of the amino acid onto the cofactor within the adenylation domain. These structures demonstrate that a conformational change within the adenylation domain guides transfer of intermediates between domains. Furthermore, one structure shows that the condensation and adenylation domains simultaneously adopt their catalytic conformations, increasing the overall efficiency in a revised structural cycle. These structures and the single-particle electron microscopy analysis demonstrate a highly dynamic domain architecture and provide the foundation for understanding the structural mechanisms that could enable engineering of novel non-ribosomal peptide synthetases.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 235-238 |
| Number of pages | 4 |
| Journal | Nature |
| Volume | 529 |
| Issue number | 7585 |
| DOIs | |
| State | Published - Jan 13 2016 |
Bibliographical note
Funding Information:Acknowledgements We thank R. Sanishvili for assistance with data collection. This work was funded in part by National Institutes of Health GM-068440 (to A.M.G.) and GM-115601 (to G.S.), and Award W81XWH-11-2-0218 from the Telemedicine and Advanced Technology Research Center of the US Army Medical Research and Materiel Command (A.M.G.). Data were collected at the GM/CA beamline of the Advanced Photon Source, which is funded by the National Cancer Institute (ACB-12002) and the National Institute of General Medical Sciences (AGM-12006) under Department of Energy contract number DE-AC02-06CH11357 to A.P.S. A Stafford Fellowship (to B.R.M.) and support from the Hauptman-Woodward Institute is acknowledged.
Funding Information:
We thank R. Sanishvili for assistance with data collection. This work was funded in part by National Institutes of Health GM-068440 (to A.M.G.) and GM-115601 (to G.S.), and Award W81XWH-11-2-0218 from the Telemedicine and Advanced Technology Research Center of the US Army Medical Research and Materiel Command (A.M.G.). Data were collected at the GM/CA beamline of the Advanced Photon Source, which is funded by the National Cancer Institute (ACB-12002) and the National Institute of General Medical Sciences (AGM-12006) under Department of Energy contract number DE-AC02-06CH11357 to A.P.S. A Stafford Fellowship (to B.R.M.) and support from the Hauptman-Woodward Institute is acknowledged.