Structures of Pathogenic Fungal FKBP12s Reveal Possible Self-Catalysis Function
Abstract
Invasive fungal infections remain difficult to treat and require novel targeting strategies. The 12-kDa FK506-binding protein (FKBP12) is a ubiquitously expressed peptidyl-prolyl isomerase with considerable homology between fungal pathogens and is thus a prime candidate for future targeting efforts to generate a panfungal strategy. Despite decades of research on FKBPs, their substrates and mechanisms of action remain unclear. Here we describe structural, biochemical, and in vivo analyses of FKBP12s from the pathogenic fungi Candida albicans, Candida glabrata, and Aspergillus fumigatus. Strikingly, multiple apo A. fumigatus and C. albicans FKBP12 crystal structures revealed a symmetric, intermolecular interaction involving the deep insertion of an active-site loop proline into the active-site pocket of an adjacent subunit. Such interactions have not been observed in previous FKBP structures. This finding indicates the possibility that this is a self-substrate interaction unique to the A. fumigatus and C. albicans fungal proteins that contain this central proline. Structures obtained with the proline in the cis and trans states provide more data in support of self-catalysis. Moreover, cysteine cross-linking experiments captured the interacting dimer, supporting the idea that it forms in solution. Finally, genetic studies exploring the impact of mutations altering the central proline and an adjacent residue providemore »
- Authors:
-
- Duke Univ. School of Medicine, Durham, NC (United States). Dept. of Biochemistry
- Duke Univ. School of Medicine, Durham, NC (United States). Division of Pediatric Infectious Diseases. Dept. of Pediatrics
- Duke Univ. School of Medicine, Durham, NC (United States). Dept. of Molecular Genetics and Microbiology
- Duke Univ. School of Medicine, Durham, NC (United States). Dept. of Biochemistry; Duke Univ. School of Medicine, Durham, NC (United States). Dept. of Radiology
- Duke Univ. School of Medicine, Durham, NC (United States). Division of Pediatric Infectious Diseases. Dept. of Pediatrics; Duke Univ. School of Medicine, Durham, NC (United States). Dept. of Molecular Genetics and Microbiology
- Publication Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1626109
- Grant/Contract Number:
- AC02-06CH11357
- Resource Type:
- Accepted Manuscript
- Journal Name:
- mBio (Online)
- Additional Journal Information:
- Journal Name: mBio (Online); Journal Volume: 7; Journal Issue: 2; Journal ID: ISSN 2150-7511
- Publisher:
- American Society for Microbiology (ASM)
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Microbiology
Citation Formats
Tonthat, Nam K., Juvvadi, Praveen Rao, Zhang, Hengshan, Lee, Soo Chan, Venters, Ron, Spicer, Leonard, Steinbach, William J., Heitman, Joseph, and Schumacher, Maria A. Structures of Pathogenic Fungal FKBP12s Reveal Possible Self-Catalysis Function. United States: N. p., 2016.
Web. doi:10.1128/mbio.00492-16.
Tonthat, Nam K., Juvvadi, Praveen Rao, Zhang, Hengshan, Lee, Soo Chan, Venters, Ron, Spicer, Leonard, Steinbach, William J., Heitman, Joseph, & Schumacher, Maria A. Structures of Pathogenic Fungal FKBP12s Reveal Possible Self-Catalysis Function. United States. https://doi.org/10.1128/mbio.00492-16
Tonthat, Nam K., Juvvadi, Praveen Rao, Zhang, Hengshan, Lee, Soo Chan, Venters, Ron, Spicer, Leonard, Steinbach, William J., Heitman, Joseph, and Schumacher, Maria A. Tue .
"Structures of Pathogenic Fungal FKBP12s Reveal Possible Self-Catalysis Function". United States. https://doi.org/10.1128/mbio.00492-16. https://www.osti.gov/servlets/purl/1626109.
@article{osti_1626109,
title = {Structures of Pathogenic Fungal FKBP12s Reveal Possible Self-Catalysis Function},
author = {Tonthat, Nam K. and Juvvadi, Praveen Rao and Zhang, Hengshan and Lee, Soo Chan and Venters, Ron and Spicer, Leonard and Steinbach, William J. and Heitman, Joseph and Schumacher, Maria A.},
abstractNote = {Invasive fungal infections remain difficult to treat and require novel targeting strategies. The 12-kDa FK506-binding protein (FKBP12) is a ubiquitously expressed peptidyl-prolyl isomerase with considerable homology between fungal pathogens and is thus a prime candidate for future targeting efforts to generate a panfungal strategy. Despite decades of research on FKBPs, their substrates and mechanisms of action remain unclear. Here we describe structural, biochemical, and in vivo analyses of FKBP12s from the pathogenic fungi Candida albicans, Candida glabrata, and Aspergillus fumigatus. Strikingly, multiple apo A. fumigatus and C. albicans FKBP12 crystal structures revealed a symmetric, intermolecular interaction involving the deep insertion of an active-site loop proline into the active-site pocket of an adjacent subunit. Such interactions have not been observed in previous FKBP structures. This finding indicates the possibility that this is a self-substrate interaction unique to the A. fumigatus and C. albicans fungal proteins that contain this central proline. Structures obtained with the proline in the cis and trans states provide more data in support of self-catalysis. Moreover, cysteine cross-linking experiments captured the interacting dimer, supporting the idea that it forms in solution. Finally, genetic studies exploring the impact of mutations altering the central proline and an adjacent residue provide evidence that any dimeric state formed in vivo, where FKBP12 concentrations are low, is transient. Taken together, these findings suggest a unique mechanism of self-substrate regulation by fungal FKBP12s, lending further novel understanding of this protein for future drug-targeting efforts.},
doi = {10.1128/mbio.00492-16},
journal = {mBio (Online)},
number = 2,
volume = 7,
place = {United States},
year = {Tue Apr 26 00:00:00 EDT 2016},
month = {Tue Apr 26 00:00:00 EDT 2016}
}
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Works referencing / citing this record:
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Figures / Tables found in this record: