title: Structures of two distinct conformations of holo-non-ribosomal peptide synthetases.
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.
authors.affiliation: Hauptman-Woodward Medical Research Institute, 700 Ellicott Street, Buffalo, New York 14203, USA.|Department of Structural Biology, University at Buffalo, Buffalo, New York 14203, USA.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Drake
authors.affiliation: Hauptman-Woodward Medical Research Institute, 700 Ellicott Street, Buffalo, New York 14203, USA.|Department of Structural Biology, University at Buffalo, Buffalo, New York 14203, USA.
authors.forename: Bradley R
authors.initials: BR
authors.lastname: Miller
authors.affiliation: Center for Drug Design and Department of Medicinal Chemistry, University of Minnesota, Minneapolis, Minnesota 55455, USA.
authors.forename: Ce
authors.initials: C
authors.lastname: Shi
authors.affiliation: Life Sciences Institute and Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Jeffrey T
authors.initials: JT
authors.lastname: Tarrasch
authors.affiliation: Hauptman-Woodward Medical Research Institute, 700 Ellicott Street, Buffalo, New York 14203, USA.|Department of Structural Biology, University at Buffalo, Buffalo, New York 14203, USA.
authors.forename: Jesse A
authors.initials: JA
authors.lastname: Sundlov
authors.affiliation: Hauptman-Woodward Medical Research Institute, 700 Ellicott Street, Buffalo, New York 14203, USA.
authors.forename: C Leigh
authors.initials: CL
authors.lastname: Allen
authors.affiliation: Life Sciences Institute and Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Georgios
authors.initials: G
authors.lastname: Skiniotis
authors.affiliation: Center for Drug Design and Department of Medicinal Chemistry, University of Minnesota, Minneapolis, Minnesota 55455, USA.
authors.forename: Courtney C
authors.initials: CC
authors.lastname: Aldrich
authors.affiliation: Hauptman-Woodward Medical Research Institute, 700 Ellicott Street, Buffalo, New York 14203, USA.|Department of Structural Biology, University at Buffalo, Buffalo, New York 14203, USA.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Gulick
mesh_terms: D040981:Acinetobacter baumannii / Q000201:enzymology*; D055162:Biocatalysis; D002352:Carrier Proteins / Q000378:metabolism; D003067:Coenzymes / Q000378:metabolism; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology*; D020035:Holoenzymes / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D010204:Pantetheine / Q000031:analogs & derivatives / Q000378:metabolism; D010453:Peptide Synthases / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary
chemical_list: D002352:Carrier Proteins; D003067:Coenzymes; D020035:Holoenzymes; D010204:Pantetheine; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase; C026269:2,3-dihydroxybenzoate - serine ligase; C003129:4'-phosphopantetheine