Dr. David J. Singel, Department of Chemistry and Biochemistry
Proteomic Profiling of Candida albicans Responses to Nitrosative and Oxidative Stress
The broad aim of this proposal 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, through analysis of post-translational modifications of protein thiols. The Specific Aims follow a sequence that begins (Aim 1) with validation of new Zdye reagents, used in conjunction with isolated model proteins, for analysis of chemical modifications of thiol groups. Zdyes promise a substantial increase in sensitivity owing to enhanced fluorescence and solubility as compared to current proteomics molecular probes. Subsequent to the physical characterization of these new dyes, the aims are to apply the technology to the analysis of the proteome of C. Albicans subjected to nitrosative and oxidative stress (Aims 2), then to identify pathways of stress counter-measures via systems modeling tested by genetic knock-outs (Aims 3). Work on the later aims is scheduled to proceed in the later years of the grant.