PDBsum entry 1frq

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Oxidoreductase PDB id
Protein chain
296 a.a. *
Waters ×221
* Residue conservation analysis
PDB id:
Name: Oxidoreductase
Title: Ferredoxin:nadp+ oxidoreductase (ferredoxin reductase) mutant e312a
Structure: Protein (ferredoxin:nadp+ oxidoreductase). Chain: a. Engineered: yes. Mutation: yes
Source: Spinacia oleracea. Spinach. Organism_taxid: 3562. Organ: leaf. Organelle: chloroplast. Expressed in: escherichia coli. Expression_system_taxid: 562.
1.95Å     R-factor:   0.190    
Authors: A.Aliverti,Z.Deng,D.Ravasi,L.Piubelli,P.A.Karplus,G.Zanetti
Key ref:
A.Aliverti et al. (1998). Probing the function of the invariant glutamyl residue 312 in spinach ferredoxin-NADP+ reductase. J Biol Chem, 273, 34008-34015. PubMed id: 9852055 DOI: 10.1074/jbc.273.51.34008
10-Oct-98     Release date:   14-Oct-98    
Go to PROCHECK summary

Protein chain
Pfam   ArchSchema ?
P00455  (FENR_SPIOL) -  Ferredoxin--NADP reductase, chloroplastic
369 a.a.
296 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Enzyme reactions 
   Enzyme class: E.C.  - Ferredoxin--NADP(+) reductase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]

Methionine Synthase
      Reaction: 2 reduced ferredoxin + NADP+ + H+ = 2 oxidized ferredoxin + NADPH
2 × reduced ferredoxin
+ NADP(+)
+ H(+)
= 2 × oxidized ferredoxin
      Cofactor: FAD
Bound ligand (Het Group name = FAD) corresponds exactly
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     oxidation-reduction process   1 term 
  Biochemical function     oxidoreductase activity     1 term  


DOI no: 10.1074/jbc.273.51.34008 J Biol Chem 273:34008-34015 (1998)
PubMed id: 9852055  
Probing the function of the invariant glutamyl residue 312 in spinach ferredoxin-NADP+ reductase.
A.Aliverti, Z.Deng, D.Ravasi, L.Piubelli, P.A.Karplus, G.Zanetti.
Ferredoxin-NADP+ reductase, the prototype of a large family of structurally related flavoenzymes, pairs single electrons carried by ferredoxin I and transfers them as a hydride to NADP+. Four mutants of the enzyme, in which Glu-312 was replaced with Asp, Gln, Leu, and Ala to probe the role of the residue charge, size, and polarity in the enzyme activity, have been heterologously expressed, purified, and characterized through steady-state, rapid kinetic studies, ligand-binding experiments, and three-dimensional structure determination by x-ray crystallography. The E312L mutant was the only one that was almost inactive (approximately 1%), whereas unexpectedly the E312A reductase was 10-100% active with the various acceptors tested. Rapid kinetic absorption spectroscopy studies demonstrated that flavin reduction by NADPH was impaired in the mutants. Furthermore, NADP(H) binding was partially perturbed. These functional and structural studies lead us to conclude that Glu-312 does not fulfil the role of proton donor during catalysis, but it is required for proper binding of the nicotinamide ring of NADP(H). In addition, its charge modulates the two one-electron redox potentials of the flavin to stabilize the semiquinone form.
  Selected figure(s)  
Figure 1.
Fig. 1. Overlay of the active-site structures of wild-type and mutant FNRs. Wild-type FNR (thick bonds), FNR-E312A (thin bonds), FNR-E312L (broken bonds), and FNR-E312Q (dotted bonds) are shown.
Figure 4.
Fig. 4. Photoreduction of the wild-type and mutant FNRs in the presence of an equimolar amount of NADP+: relationship between NADP+ and FAD reduction. circle , wild-type FNR; , FNR-E312L; , FNR-E312Q; , FNR-E312A; , FNR-E312D. Absorbance at 340 nm, which is practically an isosbestic point for the various reduction forms of FNR (24), is used to monitor NADPH formation. At each time of irradiation, the A[340] as a fraction of the maximal A[340] was plotted against the ratio of fully reduced enzyme FAD to the sum of fully reduced and oxidized enzyme flavin. The data were corrected for the presence of the FAD semiquinone.
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (1998, 273, 34008-34015) copyright 1998.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
19523113 E.Balconi, A.Pennati, D.Crobu, V.Pandini, R.Cerutti, G.Zanetti, and A.Aliverti (2009).
The ferredoxin-NADP+ reductase/ferredoxin electron transfer system of Plasmodium falciparum.
  FEBS J, 276, 3825-3836.  
19583765 M.Medina (2009).
Structural and mechanistic aspects of flavoproteins: photosynthetic electron transfer from photosystem I to NADP+.
  FEBS J, 276, 3942-3958.  
18279389 M.A.Musumeci, A.K.Arakaki, D.V.Rial, D.L.Catalano-Dupuy, and E.A.Ceccarelli (2008).
Modulation of the enzymatic efficiency of ferredoxin-NADP(H) reductase by the amino acid volume around the catalytic site.
  FEBS J, 275, 1350-1366.  
18487202 R.P.Ilagan, M.Tiso, D.W.Konas, C.Hemann, D.Durra, R.Hille, and D.J.Stuehr (2008).
Differences in a conformational equilibrium distinguish catalysis by the endothelial and neuronal nitric-oxide synthase flavoproteins.
  J Biol Chem, 283, 19603-19615.  
17635583 Rosa, A.Pennati, V.Pandini, E.Monzani, G.Zanetti, and A.Aliverti (2007).
Enzymatic oxidation of NADP+ to its 4-oxo derivative is a side-reaction displayed only by the adrenodoxin reductase type of ferredoxin-NADP+ reductases.
  FEBS J, 274, 3998-4007.  
17658942 S.Ojha, E.C.Meng, and P.C.Babbitt (2007).
Evolution of Function in the "Two Dinucleotide Binding Domains" Flavoproteins.
  PLoS Comput Biol, 3, e121.  
16755326 G.Forti, A.Agostiano, R.Barbato, R.Bassi, E.Brugnoli, G.Finazzi, F.M.Garlaschi, R.C.Jennings, B.A.Melandri, M.Trotta, G.Venturoli, G.Zanetti, D.Zannoni, and G.Zucchelli (2006).
Photosynthesis research in Italy: a review.
  Photosynth Res, 88, 211-240.  
15560800 D.L.Dupuy, D.V.Rial, and E.A.Ceccarelli (2004).
Inhibition of pea ferredoxin-NADP(H) reductase by Zn-ferrocyanide.
  Eur J Biochem, 271, 4582-4593.  
12709048 N.Carrillo, and E.A.Ceccarelli (2003).
Open questions in ferredoxin-NADP+ reductase catalytic mechanism.
  Eur J Biochem, 270, 1900-1915.  
12047373 M.Faro, C.Gómez-Moreno, M.Stankovich, and M.Medina (2002).
Role of critical charged residues in reduction potential modulation of ferredoxin-NADP+ reductase.
  Eur J Biochem, 269, 2656-2661.  
10651039 T.Mayoral, M.Medina, J.Sanz-Aparicio, C.Gómez-Moreno, and J.A.Hermoso (2000).
Structural basis of the catalytic role of Glu301 in Anabaena PCC 7119 ferredoxin-NADP+ reductase revealed by x-ray crystallography.
  Proteins, 38, 60-69.
PDB code: 1b2r
  10452605 J.K.Hurley, J.T.Hazzard, M.Martínez-Júlvez, M.Medina, C.Gómez-Moreno, and G.Tollin (1999).
Electrostatic forces involved in orienting Anabaena ferredoxin during binding to Anabaena ferredoxin:NADP+ reductase: site-specific mutagenesis, transient kinetic measurements, and electrostatic surface potentials.
  Protein Sci, 8, 1614-1622.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB code is shown on the right.