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Updated: Jun 9, 2026
Candida albicans Biofilm Chip CaBChip for High-throughput Antifungal Drug Screening
Published on: July 18, 2012
Chemoproteomic Profiling of C. albicans for Characterization of Anti-fungal Kinase Inhibitors
David J Shirley1, Meganathan Nandakumar1, Aurora Cabrera2,3
1Structural Genomics Consortium and Division of Chemical Biology and Medicinal Chemistry, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Abstract:
Candida albicans is a growing health concern as the leading causal agent of systemic candidiasis, a life-threatening fungal infection with a mortality rate of ~40% despite best available therapy. Yck2, a fungal casein kinase 1 (CK1) family member, is the cellular target of inhibitors YK-I-02 (YK) and MN-I-157 (MN). Here, multiplexed inhibitor beads paired with mass spectrometry (MIB/MS) employing ATP-competitive kinase inhibitors were used to define the selectivity of these Yck2 inhibitors across the global C. albicans proteome. The MIB matrix captured 89% of the known and predicted C. albicans protein kinases present in cell lysate. In MIB/MS competition assays, YK and MN demonstrated exquisite selectivity across the C. albicans fungal kinome with target engagement of only three CK1 homologs (Yck2, Yck22, and Hrr25) and a homolog of human p38α (Hog1). Additional chemoproteomics using a custom MN-kinobead identified only one additional C. albicans protein, confirming its remarkable fungal proteome-wide selectivity. To identify new Yck2 inhibitors with selectivity over Hog1, thirteen human CK1 kinase inhibitors were profiled for fungal kinase-binding activity using MIB/MS competition assays and in-cell NanoBRET target engagement assays. A new chemotype of family-selective Yck2 inhibitors with antifungal activity was identified. Together, these findings expand the application of MIB/MS proteomic profiling for non-human kinomes and demonstrate its utility in the discovery and development of selective inhibitors of fungal kinases with potential antimicrobial activity.
Insights
Researchers identified selective inhibitors for Candida albicans Yck2 kinase, a target for life-threatening fungal infections. This discovery advances antifungal drug development using advanced proteomic techniques.
Area of Science:
- Biochemistry
- Mycology
- Pharmacology
Background:
- Systemic candidiasis, caused by *Candida albicans*, is a life-threatening fungal infection with high mortality.
- Yck2, a fungal casein kinase 1 (CK1) family member, is a cellular target for antifungal inhibitors YK-I-02 (YK) and MN-I-157 (MN).
Purpose of the Study:
- To define the selectivity of YK and MN inhibitors across the *C. albicans* proteome using multiplexed inhibitor beads paired with mass spectrometry (MIB/MS).
- To discover new Yck2 inhibitors with selectivity over Hog1, a homolog of human p38α.
Main Methods:
- Utilized MIB/MS to profile kinase selectivity of YK and MN inhibitors against the *C. albicans* kinome.
- Employed additional chemoproteomics with a custom MN-kinobead for enhanced target identification.
- Screened thirteen human CK1 kinase inhibitors for fungal kinase-binding activity using MIB/MS and in-cell NanoBRET assays.
Main Results:
- MIB/MS captured 89% of *C. albicans* protein kinases.
- YK and MN demonstrated high selectivity, engaging only three CK1 homologs (Yck2, Yck22, Hrr25) and Hog1.
- A new chemotype of Yck2 inhibitors with antifungal activity and selectivity was identified.
Conclusions:
- MIB/MS proteomic profiling is effective for non-human kinomes, aiding in antifungal drug discovery.
- The identified selective inhibitors offer potential for developing novel antimicrobial therapies against *Candida albicans*.
