Expression, purification, and molecular characterization of Plasmodium falciparum FK506-binding protein 35 (PfFKBP35)

Protein Expr Purif. 2007 May;53(1):179-85. doi: 10.1016/j.pep.2006.12.019. Epub 2006 Dec 30.


The immunosuppressive drug FK506 binds its targets FK506-binding protein (FKBP) family and modulates cellular processes. Recent studies demonstrated that FK506 shows anti-malaria effects. Newly identified FK506-binding protein 35 from Plasmodium falciparum (PfFKBP35) is assumed to be the molecular target of FK506 in the parasite. Currently, molecular and structural basis of growth inhibition of the parasite by FK506 remains unclear. In this study, to examine characteristics of PfFKBP35 and also understand its molecular mechanism of the inhibition by FK506, we have cloned, expressed, and purified the full-length PfFKBP35 and its FK506-binding domain (FKBD). We demonstrate that the full-length PfFKBP35 and the FKBD were properly folded, and suitable for biochemical and biophysical studies. PfFKBP35 showed a basal activity in inhibiting the phosphatase activity of calcineurin in the absence of FK506, but the presence of FK506 greatly enhanced its calcineurin-inhibitory activity. Our NMR data indicate that the FKBD binds FK506 with a high affinity.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Sequence
  • Amino Acids, Aromatic / chemistry
  • Animals
  • Calcineurin / analysis
  • Calcineurin Inhibitors
  • Chromatography, Gel
  • Cloning, Molecular
  • DNA, Protozoan
  • Databases, Protein
  • Escherichia coli / genetics
  • Gene Expression
  • Genetic Vectors
  • Genome, Protozoan
  • Hydrogen Bonding
  • Molecular Sequence Data
  • Molecular Weight
  • Nuclear Magnetic Resonance, Biomolecular
  • Plasmodium falciparum / genetics
  • Plasmodium falciparum / metabolism*
  • Protein Folding
  • Protein Structure, Tertiary
  • Protozoan Proteins / chemistry
  • Protozoan Proteins / genetics
  • Protozoan Proteins / isolation & purification*
  • Protozoan Proteins / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Tacrolimus Binding Proteins / antagonists & inhibitors
  • Tacrolimus Binding Proteins / chemistry
  • Tacrolimus Binding Proteins / genetics
  • Tacrolimus Binding Proteins / isolation & purification*
  • Tacrolimus Binding Proteins / metabolism*
  • Transformation, Genetic


  • Amino Acids, Aromatic
  • Calcineurin Inhibitors
  • DNA, Protozoan
  • Protozoan Proteins
  • Recombinant Proteins
  • Calcineurin
  • Tacrolimus Binding Proteins