Structures of a Nonribosomal Peptide Synthetase Module Bound to MbtH-like Proteins Support a Highly Dynamic Domain Architecture

• Published in: The Journal of biological chemistry Vol. 291; no. 43; pp. 22559 - 22571
• Main Authors: Miller, Bradley R., Drake, Eric J., Shi, Ce, Aldrich, Courtney C., Gulick, Andrew M.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 21.10.2016; American Society for Biochemistry and Molecular Biology
• Subjects: acyl carrier protein (ACP); bacterial metabolism; BASIC BIOLOGICAL SCIENCES; enzyme mechanism; enzyme mutation; Escherichia coli - enzymology; Escherichia coli - genetics; multifunctional protein; natural product biosynthesis; Peptide Synthases - chemistry; Peptide Synthases - genetics; Peptide Synthases - metabolism; Protein Domains; Protein Structure and Folding; Pseudomonas aeruginosa - enzymology; Pseudomonas aeruginosa - genetics; siderophore; structural biology; natural product biosynthesis; acyl carrier protein (ACP); bacterial metabolism; multifunctional protein; structural biology; enzyme mechanism; enzyme mutation; siderophore
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M116.746297

Nonribosomal peptide synthetases (NRPSs) produce a wide variety of peptide natural products. During synthesis, the multidomain NRPSs act as an assembly line, passing the growing product from one module to the next. Each module generally consists of an integrated peptidyl carrier protein, an amino acid-loading adenylation domain, and a condensation domain that catalyzes peptide bond formation. Some adenylation domains interact with small partner proteins called MbtH-like proteins (MLPs) that enhance solubility or activity. A structure of an MLP bound to an adenylation domain has been previously reported using a truncated adenylation domain, precluding any insight that might be derived from understanding the influence of the MLP on the intact adenylation domain or on the dynamics of the entire NRPS module. Here, we present the structures of the full-length NRPS EntF bound to the MLPs from Escherichia coli and Pseudomonas aeruginosa. These new structures, along with biochemical and bioinformatics support, further elaborate the residues that define the MLP-adenylation domain interface. Additionally, the structures highlight the dynamic behavior of NRPS modules, including the module core formed by the adenylation and condensation domains as well as the orientation of the mobile thioesterase domain.