Infectious disease is a global threat to human health, thus defining an urgent need to develop novel antimicrobial strategies that limit the impact of life-threatening bacterial infections. The parent project supports our fundamental studies of bacterial transition metal (manganese, copper and zinc) homeostasis, sulfur metabolism and sulfide homeostasis to accelerate the pace of discovery of new antibacterial targets. In this proposal, we seek to obtain instrumentation that will allow us to probe how metal trafficking proteins, or metallochaperones, allow a bacterium to adapt to ?remodeling? of transition metal bioavailability under culture conditions that mimic host-mediated metal starvation, or ?nutritional immunity.? We propose to leverage our emerging expertise in the proteomics of the Staphylococcus aureus and Enterococcus faecalis developed under the parent project to elucidate how currently uncharacterized, COG0523 family GTPase metallochaperones impact the metalloproteome in these organisms. Partial funding is requested to acquire an integrated Perkin-Elmer NexION 2000C inductively coupled plasma-mass spectrometry (ICP-MS)-Flexar liquid chromatography (LC) system to establish a state-of-the-art metalloproteomics workflow in my laboratory. This instrumentation will be used to replace an existing ICP-MS that is no longer serviceable, and which lacks the LC-ICP-MS integration, high sensitivity, extended dynamic range and simultaneous multi-element ionization schemes characteristic of the new instrumentation, and required by this workflow. Use of this instrumentation will yield new insights into the cellular mechanisms of adaptation to the vertebrate host defense against invading microorganisms, fundamental to transition metal biology and future therapeutics discovery efforts.

Public Health Relevance

In this proposal, we request supplemental funding to acquire an integrated Perkin-Elmer NexION 2000C ICP-MS Flexar LC system to establish a state-of-the-art metalloproteomics (speciation) workflow in my laboratory. This instrumentation will be used to investigate the dynamics of transition metal (re)- distribution among cellular proteins under conditions of host-mediated ?nutritional immunity? against important bacterial pathogens.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Unknown (R35)
Project #
3R35GM118157-03S2
Application #
9712118
Study Section
Program Officer
Anderson, Vernon
Project Start
2016-06-01
Project End
2021-05-31
Budget Start
2018-06-01
Budget End
2019-05-31
Support Year
3
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Indiana University Bloomington
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
006046700
City
Bloomington
State
IN
Country
United States
Zip Code
47401
Shen, Jiangchuan; Walsh, Brenna J C; Flores-Mireles, Ana Lidia et al. (2018) Hydrogen Sulfide Sensing through Reactive Sulfur Species (RSS) and Nitroxyl (HNO) in Enterococcus faecalis. ACS Chem Biol 13:1610-1620
Glauninger, Hendrik; Zhang, Yifan; Higgins, Khadine A et al. (2018) Metal-dependent allosteric activation and inhibition on the same molecular scaffold: theĀ copper sensor CopY from Streptococcus pneumoniae. Chem Sci 9:105-118
Capdevila, Daiana A; Huerta, Fidel; Edmonds, Katherine A et al. (2018) Tuning site-specific dynamics to drive allosteric activation in a pneumococcal zinc uptake regulator. Elife 7:
Peng, Hui; Zhang, Yixiang; Palmer, Lauren D et al. (2017) Hydrogen Sulfide and Reactive Sulfur Species Impact Proteome S-Sulfhydration and Global Virulence Regulation in Staphylococcus aureus. ACS Infect Dis 3:744-755
Capdevila, Daiana A; Braymer, Joseph J; Edmonds, Katherine A et al. (2017) Entropy redistribution controls allostery in a metalloregulatory protein. Proc Natl Acad Sci U S A 114:4424-4429
Martin, Julia E; Lisher, John P; Winkler, Malcolm E et al. (2017) Perturbation of manganese metabolism disrupts cell division in Streptococcus pneumoniae. Mol Microbiol 104:334-348
Capdevila, Daiana A; Edmonds, Katherine A; Giedroc, David P (2017) Metallochaperones and metalloregulation in bacteria. Essays Biochem 61:177-200
Martin, Julia E; Edmonds, Katherine A; Bruce, Kevin E et al. (2017) The zinc efflux activator SczA protects Streptococcus pneumoniae serotype 2 D39 from intracellular zinc toxicity. Mol Microbiol 104:636-651
Shimizu, Takayuki; Shen, Jiangchuan; Fang, Mingxu et al. (2017) Sulfide-responsive transcriptional repressor SqrR functions as a master regulator of sulfide-dependent photosynthesis. Proc Natl Acad Sci U S A 114:2355-2360
Peng, Hui; Shen, Jiangchuan; Edmonds, Katherine A et al. (2017) Sulfide Homeostasis and Nitroxyl Intersect via Formation of Reactive Sulfur Species in Staphylococcus aureus. mSphere 2:

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