ArticleProceedings of the National Academy of Sciences of the United States of America2006
A protein interaction surface in nonribosomal peptide synthesis mapped by combinatorial mutagenesis and selection.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers.
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Who cites it
32 citing papers in PubMed, 67 citations in OpenAlex.
- Photoreactive Pantetheinyl Mimics Reveal Design Limitations for Crypto-Acyl Carrier Proteins.Chembiochem : a European journal of chemical biology · 2026Article
- The enterobactin biosynthetic intermediate 2,3-dihydroxybenzoic acid is a competitive inhibitor of the Escherichia coli isochorismatase EntB.Protein science : a publication of the Protein Society · 2025Article
- Evidence of isochorismate channeling between the Escherichia coli enterobactin biosynthetic enzymes EntC and EntB.Protein science : a publication of the Protein Society · 2024Article
- The specificity of intermodular recognition in a prototypical nonribosomal peptide synthetase depends on an adaptor domain.Science advances · 2024Article
- FRET Monitoring of a Nonribosomal Peptide Synthetase Elongation Module Reveals Carrier Protein Shuttling between Catalytic Domains.Angewandte Chemie (International ed. in English) · 2022Article
- Nonribosomal peptide synthetases and their biotechnological potential in Penicillium rubens.Journal of industrial microbiology & biotechnology · 2021Review
- Comparative structure, dynamics and evolution of acyl-carrier proteins from Borrelia burgdorferi, Brucella melitensis and Rickettsia prowazekii.The Biochemical journal · 2020Article
- Directed Evolution Reveals the Functional Sequence Space of an Adenylation Domain Specificity Code.ACS chemical biology · 2019Article
- Synthetic cycle of the initiation module of a formylating nonribosomal peptide synthetase.Nature · 2016Article
- Structural Biology of Nonribosomal Peptide Synthetases.Methods in molecular biology (Clifton, N.J.) · 2016Article
- Solution Structure of a Nonribosomal Peptide Synthetase Carrier Protein Loaded with Its Substrate Reveals Transient, Well-Defined Contacts.Journal of the American Chemical Society · 2015Article
- Structure and Substrate Sequestration in the Pyoluteorin Type II Peptidyl Carrier Protein PltL.Journal of the American Chemical Society · 2015Article
- Structure and catalysis in the Escherichia coli hotdog-fold thioesterase paralogs YdiI and YbdB.Biochemistry · 2014Article
- The crystal structure of BlmI as a model for nonribosomal peptide synthetase peptidyl carrier proteins.Proteins · 2014Article
- Crystal structures of the first condensation domain of CDA synthetase suggest conformational changes during the synthetic cycle of nonribosomal peptide synthetases.Journal of molecular biology · 2013Article
- Substrate placement influences reactivity in non-heme Fe(II) halogenases and hydroxylases.The Journal of biological chemistry · 2013Article
- Explorations of catalytic domains in non-ribosomal peptide synthetase enzymology.Natural product reports · 2012Review
- Structural and functional investigation of the intermolecular interaction between NRPS adenylation and carrier protein domains.Chemistry & biology · 2012Article
- Genetic and biochemical analyses of chromosome and plasmid gene homologues encoding ICL and ArCP domains in Vibrio anguillarum strain 775.Biometals : an international journal on the role of metal ions in biology, biochemistry, and medicine · 2011Article
- Structure and function of the RedJ protein, a thioesterase from the prodiginine biosynthetic pathway in Streptomyces coelicolor.The Journal of biological chemistry · 2011Article
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
Nonribosomal peptide synthetases (NRPSs) and polyketide synthases are large, multidomain enzymes that biosynthesize a number of pharmaceutically important natural products. The recognition of biosynthetic intermediates, displayed via covalent attachment to carrier proteins, by catalytic domains is critical for NRPS and polyketide synthase function. We report the use of combinatorial mutagenesis coupled with in vivo selection for the production of the Escherichia coli NRPS product enterobactin to map the surface of the aryl carrier protein (ArCP) domain of EntB that interacts with the downstream elongation module EntF. Two libraries spanning the predicted helix 2 and loop 2/helix 3 of EntB-ArCP were generated by shotgun alanine scanning and selected for their ability to support enterobactin production. From the surviving pools, we identified several hydrophobic residues (M249, F264, and A268) that were highly conserved. These residues cluster near the phosphopantetheinylated serine in a structural model, and two of these positions are in the predicted helix 3 region. Subsequent in vitro studies are consistent with the hypothesis that these residues form a surface on EntB required for interaction with EntF. These results suggest that helix 3 is a major recognition element in EntB-ArCP and demonstrate the utility of selection-based approaches for studying NRPS biosynthesis.
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