2N6Y

Solution structure of holo ArCP from yersiniabactin synthetase

Summary for 2N6Y

• Entry DOI: 10.2210/pdb2n6y/pdb
• Related: 2N6Z
• NMR Information: BMRB: 25786
• Descriptor: HMWP2 nonribosomal peptide synthetase (1 entity in total)
• Functional Keywords: carrier protein, holo, nonribosomal peptide synthetase, yersiniabactin, ligase
• Biological source: Yersinia pestis
• Total number of polymer chains: 1
• Total formula weight: 10111.40

Authors

Goodrich, A.C.,Harden, B.J.,Frueh, D.P. (deposition date: 2015-08-31, release date: 2015-09-23, Last modification date: 2015-10-07)

Primary citation

Goodrich, A.C.,Harden, B.J.,Frueh, D.P.
Solution Structure of a Nonribosomal Peptide Synthetase Carrier Protein Loaded with Its Substrate Reveals Transient, Well-Defined Contacts.
J.Am.Chem.Soc., 137:12100-12109, 2015

PubMed Abstract: Nonribosomal peptide synthetases (NRPSs) are microbial enzymes that produce a wealth of important natural products by condensing substrates in an assembly line manner. The proper sequence of substrates is obtained by tethering them to phosphopantetheinyl arms of holo carrier proteins (CPs) via a thioester bond. CPs in holo and substrate-loaded forms visit NRPS catalytic domains in a series of transient interactions. A lack of structural information on substrate-loaded carrier proteins has hindered our understanding of NRPS synthesis. Here, we present the first structure of an NRPS aryl carrier protein loaded with its substrate via a native thioester bond, together with the structure of its holo form. We also present the first quantification of NRPS CP backbone dynamics. Our results indicate that prosthetic moieties in both holo and loaded forms are in contact with the protein core, but they also sample states in which they are disordered and extend in solution. We observe that substrate loading induces a large conformational change in the phosphopantetheinyl arm, thereby modulating surfaces accessible for binding to other domains. Our results are discussed in the context of NRPS domain interactions.

PubMed: 26334259
DOI: 10.1021/jacs.5b07772

Experimental method

SOLUTION NMR