Abstract
Nonribosomal peptide synthetases (NRPSs) are modular proteins that produce peptide antibiotics and siderophores. These enzymes act as catalytic assembly lines where substrates, covalently bound to integrated carrier domains, are delivered to adjacent catalytic domains. The carrier domains are initially loaded by adenylation domains, which use two distinct conformations to catalyze sequentially the adenylation of the substrate and the thioesterification of the pantetheine cofactor. We have used a mechanism-based inhibitor to determine the crystal structure of an engineered adenylation-carrier domain protein illustrating the intermolecular interaction between the adenylation and carrier domains. This structure enabled directed mutations to improve the interaction between nonnative partner proteins. Comparison with prior NRPS adenylation domain structures provides insights into the assembly line dynamics of these modular enzymes.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 188-198 |
| Number of pages | 11 |
| Journal | Chemistry and Biology |
| Volume | 19 |
| Issue number | 2 |
| DOIs | |
| State | Published - Feb 24 2012 |
Bibliographical note
Funding Information:This work is supported in part by the National Institutes of Health (grant GM-068440 to A.M.G.). Diffraction data were collected at the Stanford Synchrotron Radiation Lightsource, a Directorate of SLAC National Accelerator Laboratory and an Office of Science User Facility operated for the U.S. Department of Energy Office of Science by Stanford University. The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research, and by the National Institutes of Health, National Center for Research Resources, Biomedical Technology Program (grant P41RR001209), and the National Institute of General Medical Sciences.