This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Leishmania are protozoan parasites that cause significant morbidity and mortality in humans, afflicting approximately 12 million people worldwide. These parasites are auxotrophic for purines and have evolved a unique set of purine transporters and salvage enzymes to scavenge these essential nutrients from their host. Hence, purine salvage components provide attractive targets for drug design. An ability to adapt to changes in the purine milieu is vital for Leishmania to survive in their host. Withdrawal of extracellular purines leads to an acute increase in key purine transporters and salvage enzymes. This augmentation is not due to an increase in mRNA abundance or stability. Since regulation is post-transcriptional, it is our goal to use proteomic methods to provide insight into the cellular networks and pathways involved in this adaptation to purine stress. Two strategies are proposed. First, to quantify the temporal changes in the Leishmania donovani proteome upon purine-starvation, an accurate mass and time (AMT) tag strategy is delineated. A broad AMT database will be constructed from parasites grown under purine-free and purine-replete conditions and used to facilitate high-throughput accurate measurements on relative protein abundance between purine-starved and purine-replete parasites. Second, since it is conjectured that part of the response to purine stress involves the post-translational modification of regulatory proteins, the phosphoproteome of purine-starved and purine-replete L. donovani will be compared. It is anticipated that these studies will be of broad impact, illuminating the means by which Leishmania adapt to changes in their host purine environment and likely providing novel targets and pathways for future therapeutic exploration.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR018522-09
Application #
8365485
Study Section
Special Emphasis Panel (ZRG1-BCMB-H (40))
Project Start
2011-07-01
Project End
2012-06-30
Budget Start
2011-07-01
Budget End
2012-06-30
Support Year
9
Fiscal Year
2011
Total Cost
$57,367
Indirect Cost
Name
Battelle Pacific Northwest Laboratories
Department
Type
DUNS #
032987476
City
Richland
State
WA
Country
United States
Zip Code
99352
Smallwood, Heather S; Duan, Susu; Morfouace, Marie et al. (2017) Targeting Metabolic Reprogramming by Influenza Infection for Therapeutic Intervention. Cell Rep 19:1640-1653
Sigdel, Tara K; Gao, Yuqian; He, Jintang et al. (2016) Mining the human urine proteome for monitoring renal transplant injury. Kidney Int 89:1244-52
Webb-Robertson, Bobbie-Jo M; Wiberg, Holli K; Matzke, Melissa M et al. (2015) Review, evaluation, and discussion of the challenges of missing value imputation for mass spectrometry-based label-free global proteomics. J Proteome Res 14:1993-2001
Ibrahim, Yehia M; Baker, Erin S; Danielson 3rd, William F et al. (2015) Development of a New Ion Mobility (Quadrupole) Time-of-Flight Mass Spectrometer. Int J Mass Spectrom 377:655-662
Ream, Thomas S; Haag, Jeremy R; Pontvianne, Frederic et al. (2015) Subunit compositions of Arabidopsis RNA polymerases I and III reveal Pol I- and Pol III-specific forms of the AC40 subunit and alternative forms of the C53 subunit. Nucleic Acids Res 43:4163-78
Malouli, Daniel; Hansen, Scott G; Nakayasu, Ernesto S et al. (2014) Cytomegalovirus pp65 limits dissemination but is dispensable for persistence. J Clin Invest 124:1928-44
Cox, Jonathan T; Marginean, Ioan; Kelly, Ryan T et al. (2014) Improving the sensitivity of mass spectrometry by using a new sheath flow electrospray emitter array at subambient pressures. J Am Soc Mass Spectrom 25:2028-37
Cao, Li; Toli?, Nikola; Qu, Yi et al. (2014) Characterization of intact N- and O-linked glycopeptides using higher energy collisional dissociation. Anal Biochem 452:96-102
Haag, Jeremy R; Brower-Toland, Brent; Krieger, Elysia K et al. (2014) Functional diversification of maize RNA polymerase IV and V subtypes via alternative catalytic subunits. Cell Rep 9:378-90
Martin, Jessica L; Yates, Phillip A; Soysa, Radika et al. (2014) Metabolic reprogramming during purine stress in the protozoan pathogen Leishmania donovani. PLoS Pathog 10:e1003938

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