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Published on: November 15, 2017
Chemoproteomic Profiling of C. albicans for Characterization of Antifungal Kinase Inhibitors
David J Shirley1, Meganathan Nandakumar1, Aurora Cabrera2,3
1Structural Genomics Consortium, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, United States.
Abstract:
Candida albicans is a major cause of systemic candidiasis, a severe fungal infection with a ∼40% mortality rate. Yck2, a casein kinase 1 (CK1) in C. albicans, is targeted by antifungal inhibitors YK-I-02 (YK) and MN-I-157 (MN). Using multiplexed inhibitor beads and mass spectrometry (MIB/MS), the selectivity of these inhibitors was determined across the fungal kinome. The MIB matrix captured 89% of C. albicans protein kinases, revealing that YK and MN selectively engage three CK1 homologues (Yck2, Yck22, and Hrr25) and a human p38α homologue (Hog1). Chemoproteomics using a custom MN-kinobead confirmed the remarkable fungal kinome selectivity. To identify new Yck2 inhibitors with selectivity over Hog1, 13 human CK1 inhibitors were screened, leading to the discovery of a new chemotype with antifungal activity. These findings highlight the utility of MIB/MS in profiling nonhuman kinomes and developing selective fungal kinase inhibitors as antimicrobial agents.
Insights
Multiplexed inhibitor beads and mass spectrometry identified new antifungal drug targets in Candida albicans. This approach aids in developing selective kinase inhibitors against fungal infections.
Area of Science:
- Biochemistry
- Mycology
- Pharmacology
Background:
- * *Candida albicans* causes systemic candidiasis, a severe fungal infection with high mortality.
- * Yck2, a casein kinase 1 (CK1) in *C. albicans*, is a target for antifungal drugs YK-I-02 (YK) and MN-I-157 (MN).
Purpose of the Study:
- * To determine the selectivity of YK and MN inhibitors across the fungal kinome.
- * To discover novel Yck2 inhibitors with selectivity over human Hog1 for antifungal drug development.
Main Methods:
- * Multiplexed inhibitor beads and mass spectrometry (MIB/MS) were used to profile the *C. albicans* kinome.
- * Chemoproteomics with a custom MN-kinobead confirmed inhibitor selectivity.
- * 13 human CK1 inhibitors were screened to identify new antifungal compounds.
Main Results:
- * MIB/MS captured 89% of *C. albicans* protein kinases.
- * YK and MN selectively targeted three CK1 homologues (Yck2, Yck22, Hrr25) and a human p38α homologue (Hog1).
- * A new chemotype with antifungal activity was discovered by screening human CK1 inhibitors.
Conclusions:
- * MIB/MS is a valuable tool for profiling nonhuman kinomes.
- * Selective fungal kinase inhibitors can be developed as antimicrobial agents.
- * The study identified potential new targets for combating *C. albicans* infections.
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