1. Structural and functional investigation of the intermolecular interaction between NRPS adenylation and carrier protein domains.
- Author
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Sundlov JA, Shi C, Wilson DJ, Aldrich CC, and Gulick AM
- Subjects
- Computer Simulation, Crystallography, X-Ray, Escherichia coli metabolism, Escherichia coli Proteins chemistry, Escherichia coli Proteins genetics, Escherichia coli Proteins metabolism, Hydrolases chemistry, Hydrolases genetics, Hydrolases metabolism, Kinetics, Ligases chemistry, Ligases genetics, Ligases metabolism, Mutation, Peptide Synthases metabolism, Protein Structure, Tertiary, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Substrate Specificity, Peptide Synthases chemistry
- 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., (Copyright © 2012 Elsevier Ltd. All rights reserved.)
- Published
- 2012
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