Crystallographic studies of the interaction between the ferredoxin-NADP+ reductase and ferredoxin from the cyanobacterium Anabaena: looking for the elusive ferredoxin molecule.

Ferredoxin-NADP(+) reductase (FNR) and its physiological electron donor ferredoxin (Fd) from the cyanobacterium Anabaena PCC7119 have been co-crystallized. The unit-cell parameters are a = b = 63.72, c = 158.02 A and the space group is P2(1)2(1)2(1). The crystal structure has been solved with 2.4 A resolution synchrotron data by molecular replacement, anomalous dispersion and R(min) search methods. For the computations, the crystal was treated as a merohedral twin. The asymmetric unit contains two FNR molecules and one ferredoxin molecule. The packing of the FNR molecules displays a nearly tetragonal symmetry (space group P4(3)2(1)2), whereas the ferredoxin arrangement is orthorhombic. This study provides the first crystallographic model of a dissociable complex between FNR and Fd.

[1]  J. Kraut,et al.  Crystal structure of a complex between electron transfer partners, cytochrome c peroxidase and cytochrome c. , 1993, Science.

[2]  A. Arakaki,et al.  Plant‐type ferredoxin‐NADP+ reductases: a basal structural framework and a multiplicity of functions , 1997, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[3]  G. Sheldrick,et al.  SHELXL: high-resolution refinement. , 1997, Methods in enzymology.

[4]  E A Merritt,et al.  Raster3D: photorealistic molecular graphics. , 1997, Methods in enzymology.

[5]  H. Kamin,et al.  Ferredoxin:NADP+ oxidoreductase. Equilibria in binary and ternary complexes with NADP+ and ferredoxin. , 1984, The Journal of biological chemistry.

[6]  J. Navaza,et al.  AMoRe: an automated package for molecular replacement , 1994 .

[7]  G. Tollin,et al.  Laser flash photolysis studies of the kinetics of reduction of ferredoxins and ferredoxin-NADP+ reductases from Anabaena PCC 7119 and spinach: electrostatic effects on intracomplex electron transfer. , 1991, Archives of biochemistry and biophysics.

[8]  Axel T. Brunger,et al.  X-PLOR Version 3.1: A System for X-ray Crystallography and NMR , 1992 .

[9]  Y. Pétillot,et al.  Refined X-ray structures of the oxidized, at 1.3 A, and reduced, at 1.17 A, [2Fe-2S] ferredoxin from the cyanobacterium Anabaena PCC7119 show redox-linked conformational changes. , 1999, Biochemistry.

[10]  A. Brunger Free R value: a novel statistical quantity for assessing the accuracy of crystal structures. , 1992 .

[11]  J. Fontecilla-Camps,et al.  X-ray structure of the ferredoxin:NADP+ reductase from the cyanobacterium Anabaena PCC 7119 at 1.8 A resolution, and crystallographic studies of NADP+ binding at 2.25 A resolution. , 1996, Journal of molecular biology.

[12]  B. Matthews Solvent content of protein crystals. , 1968, Journal of molecular biology.

[13]  J. Sancho,et al.  Interaction of ferredoxin-NADP+ reductase from Anabaena with its substrates. , 1991, Archives of biochemistry and biophysics.

[14]  G. Tollin,et al.  Structure-function relationships in Anabaena ferredoxin: correlations between X-ray crystal structures, reduction potentials, and rate constants of electron transfer to ferredoxin:NADP+ reductase for site-specific ferredoxin mutants. , 1997, Biochemistry.

[15]  C. Chothia,et al.  The atomic structure of protein-protein recognition sites. , 1999, Journal of molecular biology.

[16]  J. Pueyo,et al.  Purification of ferredoxin-NADP+ reductase, flavodoxin and ferredoxin from a single batch of the cyanobacterium Anabaena PCC 7119. , 1991, Preparative biochemistry.

[17]  T. Yeates,et al.  [22] Detecting and overcoming crystal twinning. , 1997, Methods in enzymology.

[18]  P. Kraulis A program to produce both detailed and schematic plots of protein structures , 1991 .

[19]  M G Rossmann,et al.  The molecular replacement method. , 1990, Acta crystallographica. Section A, Foundations of crystallography.

[20]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[21]  Randy J. Read,et al.  Experiences with a new translation-function program , 1987 .