4.4 Article

Structural and biochemical characterization of the cytosolic wheat cyclophilin TaCypA-1

Journal

Publisher

INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S0907444912051529

Keywords

peptidyl-prolyl cistrans isomerase; cyclophilins; Triticum aestivum

Funding

  1. Nuclear R&D Programs through the National Research Foundation (NRF) of Korea [2011-0006283]
  2. Ministry of Education, Science and Technology (MEST)
  3. 'SEED' Program of the Korea Research Council of Fundamental Science and Technology
  4. 'National Space Lab' Program through the NRF of Korea
  5. MEST [2012-0009096]
  6. KIST Institutional Program [2Z03530]
  7. KRIBB Research Initiative Program
  8. Department of Biotechnology, Ministry of Science and Technology, Government of India [BT/PR-10150/FNS/20/364/2007]
  9. JNU through 'Capacity Build-up'
  10. JNU through 'PURSE'
  11. National Research Council of Science & Technology (NST), Republic of Korea [KCM3061312, KRCF-협동-1333] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
  12. National Research Foundation of Korea [2011-0030881, 2013-PAL] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)

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Cyclophilins belong to a family of proteins that bind to the immunosuppressive drug cyclosporin A (CsA). Several members of this protein family catalyze the cistrans isomerization of peptide bonds preceding prolyl residues. The present study describes the biochemical and structural characteristics of a cytosolic cyclophilin (TaCypA-1) cloned from wheat (Triticum aestivum L.). Purified TaCypA-1 expressed in Escherichia coli showed peptidyl-prolyl cistrans isomerase activity, which was inhibited by CsA with an inhibition constant of 78.3nM. The specific activity and catalytic efficiency (kcat/Km) of the purified TaCypA-1 were 99.06 +/- 0.13nmols1mg1 and 2.32 x 105M1s1, respectively. The structures of apo TaCypA-1 and the TaCypA-1CsA complex were determined at 1.25 and 1.20 angstrom resolution, respectively, using X-ray diffraction. Binding of CsA to the active site of TaCypA-1 did not result in any significant conformational change in the apo TaCypA-1 structure. This is consistent with the crystal structure of the human cyclophilin DCsA complex reported at 0.96 angstrom resolution. The TaCypA-1 structure revealed the presence of a divergent loop of seven amino acids 48KSGKPLH54 which is a characteristic feature of plant cyclophilins. This study is the first to elucidate the structure of an enzymatically active plant cyclophilin which shows peptidyl-prolyl cistrans isomerase activity and the presence of a divergent loop.

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