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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Biochemical characterization of ferredoxin-NADP+ reductase interaction with flavodoxin in Pseudomonas putida

  • Yeom, Jin-Ki (Department of Environmental Science and Ecological Engineering, Korea University) ;
  • Park, Woo-Jun (Department of Environmental Science and Ecological Engineering, Korea University)
  • Received : 2012.03.23
  • Accepted : 2012.05.21
  • Published : 2012.08.31
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Abstract

Flavodoxin (Fld) has been demonstrated to bind to ferredoxin-NADP$^+$ reductase A (FprA) in Pseudomonas putida. Two residues ($Phe^{256}$, $Lys^{259}$) of FprA are likely to be important for interacting with Fld based on homology modeling. Site-directed mutagenesis and pH-dependent enzyme kinetics were performed to further examine the role of these residues. The catalytic efficiencies of FprA-$Ala^{259}$ and FprA-$Asp^{259}$ proteins were two-fold lower than those of the wild-type FprA. Homology modeling also strongly suggested that these two residues are important for electron transfer. Thermodynamic properties such as entropy, enthalpy, and heat capacity changes of FprA-$Ala^{259}$ and FprA-$Asp^{259}$ were examined by isothermal titration calorimetry. We demonstrated, for the first time, that $Phe^{256}$ and $Lys^{259}$ are critical residues for the interaction between FprA and Fld. Van der Waals interactions and hydrogen bonding were also more important than ionic interactions for forming the FprA-Fld complex.

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References

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