Cited by 124Signaling Pathways in DiseaseAntifungal resistance and susceptibilityMicrobial Natural Products and Biosynthesis

Harnessing calcineurin-FK506-FKBP12 crystal structures from invasive fungal pathogens to develop antifungal agents

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Juvvadi, Praveen R., Fox, David, Bobay, Benjamin G., Hoy, Michael J., Gobeil, Sophie M. C., Venters, Ronald A., Chang, Zanetta, Lin, Jackie J. +14 more·2019·Nature Communications

Abstract

Calcineurin is important for fungal virulence and a potential antifungal target, but compounds targeting calcineurin, such as FK506, are immunosuppressive. Here we report the crystal structures of calcineurin catalytic (CnA) and regulatory (CnB) subunits complexed with FK506 and the FK506-binding protein (FKBP12) from human fungal pathogens (Aspergillus fumigatus, Candida albicans, Cryptococcus neoformans and Coccidioides immitis). Fungal calcineurin complexes are similar to the mammalian complex, but comparison of fungal and human FKBP12 (hFKBP12) reveals conformational differences in the 40s and 80s loops. NMR analysis, molecular dynamic simulations, and mutations of the A. fumigatus CnA/CnB-FK506-FKBP12-complex identify a Phe88 residue, not conserved in hFKBP12, as critical for binding and inhibition of fungal calcineurin. These differences enable us to develop a less immunosuppressive FK506 analog, APX879, with an acetohydrazine substitution of the C22-carbonyl of FK506. APX879 exhibits reduced immunosuppressive activity and retains broad-spectrum antifungal activity and efficacy in a murine model of invasive fungal infection.

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Crossref · 10.1038/s41467-019-12199-1

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  • The structures identified a specific residue conserved in fungal FKBP12 but not human FKBP12 that is crucial for FK506-mediated inhibition of calcineurin.

    “These structures identified a specific residue that is conserved in the fungal FKBP12 but not the human FKBP12 and is crucial for FK506-mediated inhibition of calcineurin77”
  • APX879, an FK506 analogue with an acetohydrazine substitution of the C22 carbonyl, has 70-fold lower immunosuppressive effects than FK506 while retaining antifungal activity.

    “enabling the design of APX879, an FK506 analogue with an acetohydrazine substitution of the C22 carbonyl, which has 70-fold lower immunosuppressive effects than FK506 while retaining antifungal activity77”
  • The crystal structure of the two subunits of calcineurin in a complex with FK506 and FKBP12 from four prominent fungal pathogens, including C. neoformans, has been solved.

    “Fortunately, the crystal structure of the two subunits of calcineurin in a complex with the inhibitor FK506 and the FK506-binding protein FKBP12 from four prominent fungal pathogens, including C. neoformans, has been solved.”

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Authors
Juvvadi, P. R., Fox, D., Bobay, B. G., Hoy, M. J., Gobeil, S. M. C., Venters, R. A., Chang, Z., Lin, J. J., Averette, A. F., Cole, D. C., Barrington, B. C., Wheaton, J. D., Ciofani, M., Trzoss, M., Li, X., Lee, S. C., Chen, Y.-L., Mutz, M., Spicer, L. D., Schumacher, M. A., Heitman, J., & Steinbach, W. J.
Affiliations
  • Duke University (US)
  • Beryllium (United States) (US)
  • Duke Medical Center (US)
  • Amplyx Pharmaceuticals (United States) (US)
  • The University of Texas at San Antonio (US)
  • National Taiwan University (TW)
  • Genentech (US)
Corresponding author
  • Juvvadi, Praveen R.
  • Steinbach, William J.
Venue
Nature Communications
Published
2019-09-19
Volume
10
Issue
1
Pages
4275-4275
ISSN
2041-1723 (ISSN-L)
OpenAlex
W2973294764
Cited by
116
References
56
Citations by year
2026: 10 · 2025: 11 · 2024: 14 · 2023: 14 · 2022: 17 · 2021: 26 · 2020: 22 · 2019: 1
FWCI
5.96
Type
journal article
Language
en
Open access
Locations
  • Nature Communications (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • U.S. Department of Health & Human Services | NIH | National Institute of Allergy and Infectious Diseases (R01 AI112595-04, P01 AI104533-04, R43AI098300, R43AI106235, HHSN272200700057, HHSN272201200025C, HHSN272201700059C)
  • Ministry of Trade, Industry and Energy (N0001720)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Signaling Pathways in Disease
  • Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
  • Health Sciences › Medicine › Pharmacology › Microbial Natural Products and Biosynthesis
Keywords
calcineurin, FK506, FKBP12, antifungal agents, fungal virulence, Aspergillus fumigatus, Candida albicans, Cryptococcus neoformans, Coccidioides immitis, immunosuppressive activity, invasive fungal infections, crystal structure
MeSH terms
Animals, Antifungal Agents, Aspergillosis, Aspergillus fumigatus, Binding Sites, Candida albicans, Cells, Cultured, Coccidioides, Cryptococcosis, Cryptococcus neoformans, Female, Male, Mice, Inbred A, Mice, Inbred C57BL, Tacrolimus, Crystallography, X-Ray, Calcineurin, Tacrolimus Binding Protein 1A, Mice, Drug Discovery, Molecular Dynamics Simulation, Calcineurin Inhibitors

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