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FKBP12 dimerization mutations effect FK506 binding and differentially alter calcineurin inhibition in the human pathogen Aspergillus fumigatus.

Publication ,  Journal Article
Juvvadi, PR; Bobay, BG; Gobeil, SMC; Cole, DC; Venters, RA; Heitman, J; Spicer, LD; Steinbach, WJ
Published in: Biochem Biophys Res Commun
May 21, 2020

The 12-kDa FK506-binding protein (FKBP12) is the target of the commonly used immunosuppressive drug FK506. The FKBP12-FK506 complex binds to calcineurin and inhibits its activity, leading to immunosuppression and preventing organ transplant rejection. Our recent characterization of crystal structures of FKBP12 proteins in pathogenic fungi revealed the involvement of the 80's loop residue (Pro90) in the active site pocket in self-substrate interaction providing novel evidence on FKBP12 dimerization in vivo. The 40's loop residues have also been shown to be involved in reversible dimerization of FKBP12 in the mammalian and yeast systems. To understand how FKBP12 dimerization affects FK506 binding and influences calcineurin function, we generated Aspergillus fumigatus FKBP12 mutations in the 40's and 50's loop (F37 M/L; W60V). Interestingly, the mutants exhibited variable FK506 susceptibility in vivo indicating differing dimer strengths. In comparison to the 80's loop P90G and V91C mutants, the F37 M/L and W60V mutants exhibited greater FK506 resistance, with the F37M mutation showing complete loss in calcineurin binding in vivo. Molecular dynamics and pulling simulations for each dimeric FKBP12 protein revealed a two-fold increase in dimer strength and significantly higher number of contacts for the F37M, F37L, and W60V mutations, further confirming their varying degree of impact on FK506 binding and calcineurin inhibition in vivo.

Duke Scholars

Published In

Biochem Biophys Res Commun

DOI

EISSN

1090-2104

Publication Date

May 21, 2020

Volume

526

Issue

1

Start / End Page

48 / 54

Location

United States

Related Subject Headings

  • Tacrolimus Binding Protein 1A
  • Tacrolimus
  • Protein Structure, Secondary
  • Protein Multimerization
  • Protein Binding
  • Mutation
  • Mutant Proteins
  • Humans
  • Fungal Proteins
  • Computer Simulation
 

Citation

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MLA
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Juvvadi, P. R., Bobay, B. G., Gobeil, S. M. C., Cole, D. C., Venters, R. A., Heitman, J., … Steinbach, W. J. (2020). FKBP12 dimerization mutations effect FK506 binding and differentially alter calcineurin inhibition in the human pathogen Aspergillus fumigatus. Biochem Biophys Res Commun, 526(1), 48–54. https://doi.org/10.1016/j.bbrc.2020.03.062
Juvvadi, Praveen R., Benjamin G. Bobay, Sophie M. C. Gobeil, D Christopher Cole, Ronald A. Venters, Joseph Heitman, Leonard D. Spicer, and William J. Steinbach. “FKBP12 dimerization mutations effect FK506 binding and differentially alter calcineurin inhibition in the human pathogen Aspergillus fumigatus.Biochem Biophys Res Commun 526, no. 1 (May 21, 2020): 48–54. https://doi.org/10.1016/j.bbrc.2020.03.062.
Juvvadi PR, Bobay BG, Gobeil SMC, Cole DC, Venters RA, Heitman J, et al. FKBP12 dimerization mutations effect FK506 binding and differentially alter calcineurin inhibition in the human pathogen Aspergillus fumigatus. Biochem Biophys Res Commun. 2020 May 21;526(1):48–54.
Juvvadi, Praveen R., et al. “FKBP12 dimerization mutations effect FK506 binding and differentially alter calcineurin inhibition in the human pathogen Aspergillus fumigatus.Biochem Biophys Res Commun, vol. 526, no. 1, May 2020, pp. 48–54. Pubmed, doi:10.1016/j.bbrc.2020.03.062.
Juvvadi PR, Bobay BG, Gobeil SMC, Cole DC, Venters RA, Heitman J, Spicer LD, Steinbach WJ. FKBP12 dimerization mutations effect FK506 binding and differentially alter calcineurin inhibition in the human pathogen Aspergillus fumigatus. Biochem Biophys Res Commun. 2020 May 21;526(1):48–54.
Journal cover image

Published In

Biochem Biophys Res Commun

DOI

EISSN

1090-2104

Publication Date

May 21, 2020

Volume

526

Issue

1

Start / End Page

48 / 54

Location

United States

Related Subject Headings

  • Tacrolimus Binding Protein 1A
  • Tacrolimus
  • Protein Structure, Secondary
  • Protein Multimerization
  • Protein Binding
  • Mutation
  • Mutant Proteins
  • Humans
  • Fungal Proteins
  • Computer Simulation