title: The 1.8 A crystal structure of PA2412, an MbtH-like protein from the pyoverdine cluster of Pseudomonas aeruginosa.
abstract: Many bacteria use nonribosomal peptide synthetase (NRPS) proteins to produce peptide antibiotics and siderophores. The catalytic domains of the NRPS proteins are usually linked in large multidomain proteins. Often, additional proteins are coexpressed with NRPS proteins that modify the NRPS peptide products, ensure the availability of substrate building blocks, or play a role in the import or export of the NRPS product. Many NRPS clusters include a small protein of approximately 80 amino acids with homology to the MbtH protein of mycobactin synthesis in Mycobacteria tuberculosis; no function has been assigned to these proteins. Pseudomonas aeruginosa utilizes an NRPS cluster to synthesize the siderophore pyoverdine. The pyoverdine peptide contains a dihydroxyquinoline-based chromophore, as well as two formyl-N-hydroxyornithine residues, which are involved in iron binding. The pyoverdine cluster contains four modular NRPS enzymes and 10-15 additional proteins that are essential for pyoverdine production. Coexpressed with the pyoverdine synthetic enzymes is a 72-amino acid MbtH-like family member designated PA2412. We have determined the three-dimensional structure of the PA2412 protein and describe here the structure and the location of conserved regions. Additionally, we have further analyzed a deletion mutant of the PA2412 protein for growth and pyoverdine production. Our results demonstrate that PA2412 is necessary for the production or secretion of pyoverdine at normal levels. The PA2412 deletion strain is able to use exogenously produced pyoverdine, showing that there is no defect in the uptake or utilization of the iron-pyoverdine complex.
authors.affiliation: Hauptman-Woodward Medical Research Institute, State University of New York at Buffalo, 700 Ellicott Street, Buffalo, NY 14203, USA.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Drake
authors.forename: Jin
authors.initials: J
authors.lastname: Cao
authors.forename: Jun
authors.initials: J
authors.lastname: Qu
authors.forename: Manish B
authors.initials: MB
authors.lastname: Shah
authors.forename: Robert M
authors.initials: RM
authors.lastname: Straubinger
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Gulick
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites / Q000502:physiology; D001693:Biological Transport, Active / Q000235:genetics; D018360:Crystallography, X-Ray; D017353:Gene Deletion; D007501:Iron / Q000378:metabolism; D005810:Multigene Family; D009169:Mycobacterium tuberculosis / Q000201:enzymology / Q000235:genetics; D009842:Oligopeptides / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D010080:Oxazoles / Q000737:chemistry / Q000378:metabolism; D010453:Peptide Synthases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D017385:Sequence Homology; D017262:Siderophores / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D009842:Oligopeptides; D010080:Oxazoles; D017262:Siderophores; C018608:mycobactins; C042453:pyoverdin; D007501:Iron; D010453:Peptide Synthases