Siderophores are low molecular weight iron chelators produced by bacteria to scavenge iron from the environment, and are frequently required for bacterial virulence. The long-term goal of the research in this laboratory is to provide a detailed mechanistic and structural understanding of enzymes in the siderophore biosynthetic pathways that are targets for drug discovery. Toward that end, we propose to investigate the nonribosomal peptide synthetase accessory enzymes that are involved in the production of the siderophores pyochelin and pyoverdin from the bacteria P. aeruginosa. Two of the four enzymes to be studied, the isochorismate synthase (PchA) and isochorismate-pyruvate lyase (PchB), are ideal model systems for making fundamental advances in understanding alternate reaction pathways for catalysis. The studies of the ornithine hydroxylase (PvdA) show promise for novel flavin biochemistry, and the first characterization of the hydroxyornithine transformylase (PvdF) is proposed. We will use a multifaceted approach that includes enzyme kinetic analysis, structural biology and computational enzymology. Narrative Pseudomonas aeruginosa is a dangerous pathogen that is a common cause of infections in susceptible hosts, including cancer and AIDS patients, those with immune deficiencies, cystic fibrosis patients, burn patients and other at risk individuals including infants. The goal of this work is to provide a fundamental understanding of four enzymes that promote virulence in P. aeruginosa so that this information can be exploited to generate new antimicrobial drugs.

Public Health Relevance

Pseudomonas aeruginosa is a dangerous pathogen that is a common cause of infections in susceptible hosts, including cancer and AIDS patients, those with immune deficiencies, cystic fibrosis patients, burn patients and other at risk individuals including infants. The goal of this work is to provide a fundamental understanding of four enzymes that promote virulence in P. aeruginosa so that this information can be exploited to generate new antimicrobial drugs.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Scientist Development Award - Research (K02)
Project #
5K02AI093675-04
Application #
8616713
Study Section
Microbiology and Infectious Diseases B Subcommittee (MID)
Program Officer
Taylor, Christopher E,
Project Start
2011-03-01
Project End
2015-02-28
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
4
Fiscal Year
2014
Total Cost
$106,920
Indirect Cost
$7,920
Name
University of Kansas Lawrence
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
076248616
City
Lawrence
State
KS
Country
United States
Zip Code
66045
Meneely, Kathleen M; Sundlov, Jesse A; Gulick, Andrew M et al. (2016) An Open and Shut Case: The Interaction of Magnesium with MST Enzymes. J Am Chem Soc 138:9277-93
Meneely, Kathleen M; Ronnebaum, Trey A; Riley, Andrew P et al. (2016) Holo Structure and Steady State Kinetics of the Thiazolinyl Imine Reductases for Siderophore Biosynthesis. Biochemistry 55:5423-33
Lamb, Audrey L (2015) Breaking a pathogen's iron will: Inhibiting siderophore production as an antimicrobial strategy. Biochim Biophys Acta 1854:1054-70
Lamb, Audrey L; Kappock, T Joseph; Silvaggi, Nicholas R (2015) You are lost without a map: Navigating the sea of protein structures. Biochim Biophys Acta 1854:258-68
Meneely, Kathleen M; Luo, Qianyi; Riley, Andrew P et al. (2014) Expanding the results of a high throughput screen against an isochorismate-pyruvate lyase to enzymes of a similar scaffold or mechanism. Bioorg Med Chem 22:5961-9
Chilton, Annemarie S; Ellis, Ashley L; Lamb, Audrey L (2014) Structure of an Aspergillus fumigatus old yellow enzyme (EasA) involved in ergot alkaloid biosynthesis. Acta Crystallogr F Struct Biol Commun 70:1328-32
Frederick, Rosanne E; Ojha, Sunil; Lamb, Audrey et al. (2014) How pH modulates the reactivity and selectivity of a siderophore-associated flavin monooxygenase. Biochemistry 53:2007-16
Meneely, Kathleen M; Luo, Qianyi; Lamb, Audrey L (2013) Redesign of MST enzymes to target lyase activity instead promotes mutase and dehydratase activities. Arch Biochem Biophys 539:70-80
Meneely, Kathleen M; Luo, Qianyi; Dhar, Prajnaparamita et al. (2013) Lysine221 is the general base residue of the isochorismate synthase from Pseudomonas aeruginosa (PchA) in a reaction that is diffusion limited. Arch Biochem Biophys 538:49-56
Meneely, Kathleen M; Lamb, Audrey L (2012) Two structures of a thiazolinyl imine reductase from Yersinia enterocolitica provide insight into catalysis and binding to the nonribosomal peptide synthetase module of HMWP1. Biochemistry 51:9002-13

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