Affordable Access

deepdyve-link deepdyve-link
Publisher Website

Biochemical characterization of ferredoxin-NADP(+) reductase interaction with flavodoxin in Pseudomonas putida.

Authors
  • 1
  • 1 Department of Environmental Science and Ecological Engineering, Korea University, Seoul, Korea. , (North Korea)
Type
Published Article
Journal
BMB reports
1976-670X
Publication Date
Volume
45
Issue
8
Pages
476–481
Identifiers
DOI: 10.5483/BMBRep.2012.45.8.071
PMID: 22917033
Source
Medline
License
Unknown

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. Sitedirected 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.

Statistics

Seen <100 times