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. Candida albicans is a prevalent human pathogen, which presents a serious health risk in immunocompromised individuals. The first line of host defense against C. albicans involves innate immune responses that challenge the invading pathogen with macrophage- and neutrophil-generated nitrosative and oxidative stress. The counter-measures that pathogens muster against this stress are not well understood, but appear to include roles for proteins involved in enzymatic detoxification of stress-inducing chemical agents, and in signaling cascades that regulate of cell growth. Very little is known, however, at the molecular level, of the responses of C. albicans to oxidative and nitrosative stress. The broad goal of this research is to develop enhanced differential proteomics methods and to apply them to advance our understanding of the molecular bases of C. albicans defenses against host nitrosative and oxidative challenges. Elucidation of the primary biochemical steps in defense pathways will establish new targets for the design of more effective and specific drugs against C. albicans. This research will focus on early responses that are likely to involve reversible modification of reactive protein thiols that yield S-nitrosothiols (RSNO), S- sulfenic acids (RSOH) and transient disulfices. The fundamental questions that we will address are: (1) what components of the C. albicans proteome are modified by different types and levels of oxidative and nitrosative stress;(2) which of these modifications involve redox signaling processes in which protein sensors detect and transduce the stress;and (3) what biochemical pathways are modulated by stress-responsive protein modifications? Differential thiol redox profiling (dThiRP) - featuring a new generation of """"""""Zdyes"""""""", that promise unsurpassed threshold sensitivities and the capacity for multi-color differential profiling - will be used to address these questions. Specifically, the research program entails: (1) validation of thiol """"""""switch"""""""" detection chemistry with the Zdye labels;(2) global proteomic measurements of changes in protein levels, S-nitrosothiols, sulfenic acids, and disulfides in C. albicans cells subject to exogenous chemical, and macrophage challenge;(3) measurements of the relative resistance to chemical and macrophage challenge of homozygous and heterozygous KO strains, suggested by modeling of the wild-type proteomic data.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Exploratory Grants (P20)
Project #
5P20RR024237-04
Application #
8359570
Study Section
National Center for Research Resources Initial Review Group (RIRG)
Project Start
2011-03-01
Project End
2012-02-29
Budget Start
2011-03-01
Budget End
2012-02-29
Support Year
4
Fiscal Year
2011
Total Cost
$198,165
Indirect Cost
Name
Montana State University - Bozeman
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
625447982
City
Bozeman
State
MT
Country
United States
Zip Code
59717
Folsom, James Patrick; Carlson, Ross P (2015) Physiological, biomass elemental composition and proteomic analyses of Escherichia coli ammonium-limited chemostat growth, and comparison with iron- and glucose-limited chemostat growth. Microbiology 161:1659-70
Blosser, Sara J; Merriman, Brittney; Grahl, Nora et al. (2014) Two C4-sterol methyl oxidases (Erg25) catalyse ergosterol intermediate demethylation and impact environmental stress adaptation in Aspergillus fumigatus. Microbiology 160:2492-506
Mason, Katelyn E; Tripet, Brian P; Parrott, David et al. (2014) ¹H, ¹³C, ¹⁵N backbone and side chain NMR resonance assignments for the N-terminal RNA recognition motif of the HvGR-RBP1 protein involved in the regulation of barley (Hordeum vulgare L.) senescence. Biomol NMR Assign 8:149-53
Chung, Dawoon; Thammahong, Arsa; Shepardson, Kelly M et al. (2014) Endoplasmic reticulum localized PerA is required for cell wall integrity, azole drug resistance, and virulence in Aspergillus fumigatus. Mol Microbiol 92:1279-98
Duffus, Benjamin R; Ghose, Shourjo; Peters, John W et al. (2014) Reversible H atom abstraction catalyzed by the radical S-adenosylmethionine enzyme HydG. J Am Chem Soc 136:13086-9
Heinemann, Joshua; Noon, Brigit; Mohigmi, Mohammad J et al. (2014) Real-time digitization of metabolomics patterns from a living system using mass spectrometry. J Am Soc Mass Spectrom 25:1755-62
Fonner, Brittany A; Tripet, Brian P; Lui, Mengyao et al. (2014) ¹H, ¹³C, ¹⁵N backbone and side chain NMR resonance assignments of the N-terminal NEAr iron transporter domain 1 (NEAT 1) of the hemoglobin receptor IsdB of Staphylococcus aureus. Biomol NMR Assign 8:201-5
Bernstein, Hans C; Kesaano, Maureen; Moll, Karen et al. (2014) Direct measurement and characterization of active photosynthesis zones inside wastewater remediating and biofuel producing microalgal biofilms. Bioresour Technol 156:206-15
Harvey, Emily; Heys, Jeffrey; Gedeon, Tomáš (2014) Quantifying the effects of the division of labor in metabolic pathways. J Theor Biol 360:222-42
Weaver Jr, Alan J; Shepard, Joyce B; Wilkinson, Royce A et al. (2014) Antibacterial activity of THAM Trisphenylguanide against methicillin-resistant Staphylococcus aureus. PLoS One 9:e97742

Showing the most recent 10 out of 38 publications