Abstract
Multi-drug resistance is becoming an increasingly severe clinical challenge not only among pathogenic bacteria but among fungal pathogens as well. Drug design is inherently more challenging for the eukaryotic fungi due to their closer evolutionary similarity to humans. The recent rapid expansion in invasive infections throughout the world by Candida auris is of particular concern due to a substantial mortality rate, comparatively facile transmission, and an increasing level of resistance to all three of the major classes of anti-fungal drugs. One promising avenue for the development of an alternative class of anti-fungal agents currently under investigation is for drugs against the FK506-binding protein FKBP12 which, when bound to that drug, inhibits the fungal calcineurin signaling pathway with a resultant diminution in virulence. The specific challenge to this approach is that the homologous human calcineurin pathway functions in controlling the tissue immunity response, so that drug selectivity for the fungal pathway must be designed. To facilitate such efforts, we report the nearly complete backbone and sidechain resonances for the FKBP12 proteins of both Candida auris and clinically significant Candida glabrata fungi.
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Blankenship JR, Wormley FL, Boyce MK, Schell WA, Filler SG (2003) Calcineurin is essential for Candida albicans survival in serum and virulence. Eukaryot Cell 2:422–430. https://doi.org/10.1128/EC.2.3.422-430.2003
Borst A, Raimer MT, Warnock DW, Morrison CJ, Arthington-Skaggs BA (2005) Rapid acquisition of stable azole resistance by Candida glabrata isolates obtained before the clinical introduction of fluconazole. Antimicrob Agents Chemother 49:783–787. https://doi.org/10.1128/AAC.49.2.783-787.2005
Chen YL, Lehman VN, Lewit Y, Averette AF, Heitman J (2013) Calcineurin governs thermotolerance and virulence of Cryptococcus gatti. Genetics 3:527–539. https://doi.org/10.1534/g3.112.004242
Forsberg K, Woodworth K, Walters M, Berkow EL, Jackson B, Chiller T, Vallabhaneni S (2019) Candida auris: the recent emergence of a multidrug-resistant fungal pathogen. Med Mycol 57:1–12. https://doi.org/10.1093/mmy/mmy156
Gobeil SMC, Bobay BG, Spicer LD, Venters RA (2019) 15N, 13C and 1H resonance assignments of FKBP12 proteins from the pathogenic fungi Mucor circinelloides and Aspergillus fumigatus. Biomol NMR Assign 13:207–212. https://doi.org/10.1007/s12104-019-09878-x
Juvvadi PR, Fox DI, Bobay BG, Hoy MJ, Gobeil SMC, Venters RA, Chang Z, Lin JJ, Averette AF, Cole C, Barrington BC, Wheaton JD, Ciofani M, Trzoss M, Li X, Lee SC, Chen YL, Mutz M, Spicer LD, Schumacher MA, Heitman J, Steinbach WJ (2019) Harnessing calcineurin-FK506-FKBP12 crystal structures from invasive fungal pathogens to develop antifungal agents. Nat Commun. https://doi.org/10.1038/s41467-019-12199-1
Kay LE, Xu GY, Singer AU, Muhandiram DR, Forman-Kay JD (1993) A gradient-enhanced HCCH TOCSY experiment for recording side-chain H-1 and C-13 correlations in H2O samples of proteins. J Magn Reson B 101:333–337. https://doi.org/10.1006/jmrb.1993.1053
Kay LE, Xu GY, Yamazaki T (1994) Enhanced-sensitivity triple-resonance spectroscopy with minimal H2O saturation. J Magn Reson A 109:129–133. https://doi.org/10.1006/jmra.1994.1145
Lee WG, Shin JH, Uh Y (2011) First three reported cases of nosocomial fungemia caused by Candida auris. J Clin Microbiol 49:3139–3142. https://doi.org/10.1128/JCM.00319-11
Lockhart SR, Etienne KA, Vallabhaneni S, Farooqi J, Chowdhary A, Govender NP, Colombo AL, Calvo B, Cuomo CA, Desjardins CA, Berkow EL, Castanheira M, Magobo RE, Jabeen K, Asghar RJ, Meis JF, Jackson B, Chiller T, Litvintseva AP (2017) Simultaneous emergence of multidrug-resistant Candida auris on 3 continents confirmed by whole-genome sequencing and epidemiological analyses. Clin Infect Dis 64:134–140. https://doi.org/10.1093/cid/ciw691
Lone SA, Ahmad A (2019) Candida auris—the growing menace to global health. Mycoses 62:620–637. https://doi.org/10.1111/myc.12904
Muhandiram DR, Kay LE (1994) Gradient-enhanced triple-resonance three-dimensional NMR experiments with improved sensitivity. J Magn Reson B 103:203–216. https://doi.org/10.1006/jmrb.1994.1032
Mustafi SM, Chen H, Li H, Lemaster DM, Hernández G (2013) Analyzing the visible conformational substates of the FK506-binding protein FKBP12. Biochem J 453:371–380. https://doi.org/10.1042/BJ20130276
Odom A, Muir S, Lim E, Toffaletti DL, Perfect JR, Heitman J (1997) Calcineurin is required for virulence of Cryptococcus neoformans. EMBO J 16:2576–2589. https://doi.org/10.1093/emboj/16.10.2576
Pais P, Galocha M, Viana R, Cavalheiro M, Pereira D, Teixeira MC (2019) Microevolution of the pathogenic yeasts Candida albicans and Candida glabrata during antifungal therapy and host infection. Microbial Cell 6:142–159. https://doi.org/10.15698/mic2019.03.670
Palmer AG, Cavanagh J, Wright PE, Rance M (1991) Sensitivity improvement in proton-detected two-dimensional heteronuclear correlation NMR spectroscopy. J Magn Reson 93:151–170. https://doi.org/10.1016/0022-2364(91)90036-S
Park HS, Lee SC, Cardenas ME, Heitman J (2019) Calcium-calmodulin-calcineurin signaling: a globally conserved virulence cascade in eukaryotic microbial pathogens. Cell Host Microbe 26:453–462. https://doi.org/10.1016/j.chom.2019.08.004
Rhodes J, Fisher MC (2019) Global epidemiology of emerging Candida auris. Cuur Opin Microbiol 52:84–89. https://doi.org/10.1016/j.mib.2019.05.008
Steinbach WJ, Cramer RA Jr, Perfect BZ, Asfaw YG, Sauer TC, Najvar LK, Kirkpatrick WR, Patterson TF, Benjamin DK Jr, Heitman J (2006) Calcineurin controls growth, morphology, ad pathogenicity in Aspergillus fumigatus. Eukaryot Cell 5:1091–1103. https://doi.org/10.1128/EC.00139-06
Tonthat NK, Juvvadi PR, Zhang H, Lee SC, Venters R, Spicer L, Steinbach WJ, Heitman J, Schumacher MA (2016) Structures of pathogenic fungal fkbp12s reveal possible self-catalysis function. mBio 7:e00492–00416. https://doi.org/10.1128/mBio.00492-16
Vuister GW, Bax A (1992) Resolution enhancement and spectral editing of uniformily 13C-enriched proteins by homonuclear broadband 13C decoupling. J Magn Reson 98:428–435. https://doi.org/10.1016/0022-2364(92)90144-V
Wishart DS, Bigam CG, Yao J, Abildgaard F, Dyson HJ, Oldfield E, Markley JL, Sykes BD (1995) 1H, 13C, and 15N chemical shift referencing in biomolecular NMR. J Biomol NMR 6:135–140. https://doi.org/10.1007/BF00211777
Yamazaki T, Lee W, Arrowsmith CH, Muhandiram DR, Kay LE (1994) A suite of triple resonance NMR experiments for the backbone assignment of 15N, 13C, 2H labeled proteins with high sensitivity. J Am Chem Soc 116:11655–11666. https://doi.org/10.1021/ja00105a005
Acknowledgements
We acknowledge the use of the NMR facility at the Wadsworth Center. This work was supported in part by National Institutes of Health [GM 119152 to G.H.].
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Bashir, Q., LeMaster, D.M. & Hernández, G. 1H, 13C, 15 N chemical shift assignments of the FKBP12 protein from the pathogenic fungi Candida auris and Candida glabrata. Biomol NMR Assign 14, 105–109 (2020). https://doi.org/10.1007/s12104-020-09928-9
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DOI: https://doi.org/10.1007/s12104-020-09928-9