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Scanning Quadrupole Data-Independent Acquisition, Part B: Application to the Analysis of the Calcineurin-Interacting Proteins during Treatment of Aspergillus fumigatus with Azole and Echinocandin Antifungal Drugs.

Publication ,  Journal Article
Juvvadi, PR; Moseley, MA; Hughes, CJ; Soderblom, EJ; Lennon, S; Perkins, SR; Thompson, JW; Geromanos, SJ; Wildgoose, J; Richardson, K ...
Published in: J Proteome Res
February 2, 2018

Calcineurin is a critical cell-signaling protein that orchestrates growth, stress response, virulence, and antifungal drug resistance in several fungal pathogens. Blocking calcineurin signaling increases the efficacy of several currently available antifungals and suppresses drug resistance. We demonstrate the application of a novel scanning quadrupole DIA method for the analysis of changes in the proteins coimmunoprecipitated with calcineurin during therapeutic antifungal drug treatments of the deadly human fungal pathogen Aspergillus fumigatus. Our experimental design afforded an assessment of the precision of the method as demonstrated by peptide- and protein-centric analysis from eight replicates of the study pool QC samples. Two distinct classes of clinically relevant antifungal drugs that are guideline recommended for the treatment of invasive "aspergillosis" caused by Aspergillus fumigatus, the azoles (voriconazole) and the echinocandins (caspofungin and micafungin), which specifically target the fungal plasma membrane and the fungal cell wall, respectively, were chosen to distinguish variations occurring in the proteins coimmunoprecipitated with calcineurin. Novel potential interactors were identified in response to the different drug treatments that are indicative of the possible role for calcineurin in regulating these effectors. Notably, treatment with voriconazole showed increased immunoprecipitation of key proteins involved in membrane ergosterol biosynthesis with calcineurin. In contrast, echinocandin (caspofungin or micafungin) treatments caused increased immunoprecipitation of proteins involved in cell-wall biosynthesis and septation. Furthermore, abundant coimmunoprecipitation of ribosomal proteins with calcineurin occurred exclusively in echinocandins treatment, indicating reprogramming of cellular growth mechanisms during different antifungal drug treatments. While variations in the observed calcineurin immunoprecipitated proteins may also be due to changes in their expression levels under different drug treatments, this study suggests an important role for calcineurin-dependent cellular mechanisms in response to antifungal treatment of A. fumigatus that warrants future studies.

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Published In

J Proteome Res

DOI

EISSN

1535-3907

Publication Date

February 2, 2018

Volume

17

Issue

2

Start / End Page

780 / 793

Location

United States

Related Subject Headings

  • Voriconazole
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Ribosomal Proteins
  • Protein Interaction Mapping
  • Protein Binding
  • Molecular Sequence Annotation
  • Micafungin
  • Lipopeptides
  • Immunoprecipitation
  • Green Fluorescent Proteins
 

Citation

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Juvvadi, P. R., Moseley, M. A., Hughes, C. J., Soderblom, E. J., Lennon, S., Perkins, S. R., … Steinbach, W. J. (2018). Scanning Quadrupole Data-Independent Acquisition, Part B: Application to the Analysis of the Calcineurin-Interacting Proteins during Treatment of Aspergillus fumigatus with Azole and Echinocandin Antifungal Drugs. J Proteome Res, 17(2), 780–793. https://doi.org/10.1021/acs.jproteome.7b00499
Juvvadi, Praveen R., M Arthur Moseley, Christopher J. Hughes, Erik J. Soderblom, Sarah Lennon, Simon R. Perkins, J Will Thompson, et al. “Scanning Quadrupole Data-Independent Acquisition, Part B: Application to the Analysis of the Calcineurin-Interacting Proteins during Treatment of Aspergillus fumigatus with Azole and Echinocandin Antifungal Drugs.J Proteome Res 17, no. 2 (February 2, 2018): 780–93. https://doi.org/10.1021/acs.jproteome.7b00499.
Juvvadi PR, Moseley MA, Hughes CJ, Soderblom EJ, Lennon S, Perkins SR, Thompson JW, Geromanos SJ, Wildgoose J, Richardson K, Langridge JI, Vissers JPC, Steinbach WJ. Scanning Quadrupole Data-Independent Acquisition, Part B: Application to the Analysis of the Calcineurin-Interacting Proteins during Treatment of Aspergillus fumigatus with Azole and Echinocandin Antifungal Drugs. J Proteome Res. 2018 Feb 2;17(2):780–793.
Journal cover image

Published In

J Proteome Res

DOI

EISSN

1535-3907

Publication Date

February 2, 2018

Volume

17

Issue

2

Start / End Page

780 / 793

Location

United States

Related Subject Headings

  • Voriconazole
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Ribosomal Proteins
  • Protein Interaction Mapping
  • Protein Binding
  • Molecular Sequence Annotation
  • Micafungin
  • Lipopeptides
  • Immunoprecipitation
  • Green Fluorescent Proteins