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PDBsum entry 2c7g

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protein ligands metals links
Oxidoreductase PDB id
2c7g
Jmol PyMol
Contents
Protein chain
453 a.a. *
Ligands
FAD
ODP
Metals
_NA
Waters ×630
* Residue conservation analysis
PDB id:
2c7g
Name: Oxidoreductase
Title: Fpra from mycobacterium tuberculosis: his57gln mutant
Structure: NADPH-ferredoxin reductase fpra. Chain: a. Synonym: fpra, nfr. Engineered: yes. Mutation: yes
Source: Mycobacterium tuberculosis. Organism_taxid: 1773. Expressed in: escherichia coli. Expression_system_taxid: 511693.
Biol. unit: Dimer (from PDB file)
Resolution:
1.8Å     R-factor:   0.182     R-free:   0.219
Authors: A.Pennati,A.Razeto,M.De Rosa,V.Pandini,M.A.Vanoni, A.Aliverti,A.Mattevi,A.Coda,G.Zanetti
Key ref: A.Pennati et al. (2006). Role of the His57-Glu214 ionic couple located in the active site of Mycobacterium tuberculosis FprA. Biochemistry, 45, 8712-8720. PubMed id: 16846214
Date:
24-Nov-05     Release date:   26-Jul-06    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam  
P9WIQ3  (FPRA_MYCTU) -  NADPH-ferredoxin reductase FprA
Seq:
Struc:
456 a.a.
453 a.a.*
Key:    Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Enzyme reactions 
   Enzyme class: E.C.1.18.1.2  - Ferredoxin--NADP(+) reductase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]

      Pathway:
Methionine Synthase
      Reaction: 2 reduced ferredoxin + NADP+ + H+ = 2 oxidized ferredoxin + NADPH
2 × reduced ferredoxin
+
NADP(+)
Bound ligand (Het Group name = ODP)
matches with 97.00% similarity
+ H(+)
= 2 × oxidized ferredoxin
+ NADPH
      Cofactor: FAD
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!
  Cellular component     cell wall   2 terms 
  Biological process     oxidation-reduction process   1 term 
  Biochemical function     oxidoreductase activity     5 terms  

 

 
    reference    
 
 
Biochemistry 45:8712-8720 (2006)
PubMed id: 16846214  
 
 
Role of the His57-Glu214 ionic couple located in the active site of Mycobacterium tuberculosis FprA.
A.Pennati, A.Razeto, M.de Rosa, V.Pandini, M.A.Vanoni, A.Mattevi, A.Coda, A.Aliverti, G.Zanetti.
 
  ABSTRACT  
 
Mycobacterium tuberculosis FprA is a NADPH-ferredoxin reductase, functionally and structurally similar to the mammalian adrenodoxin reductase. It is presumably involved in supplying electrons to one or more of the pathogen's cytochrome P450s through reduced ferredoxins. It has been proposed on the basis of crystallographic data (Bossi, R. T., et al. (2002) Biochemistry 41, 8807-8818) that the highly conserved His57 and Glu214 whose side chains are H-bonded are involved in catalysis. Both residues were individually changed to nonionizable amino acyl residues through site-directed mutagenesis. Steady-state kinetics showed that the role of Glu214 in catalysis is negligible. On the contrary, the substitutions of His57 markedly impaired the catalytic efficiency of FprA for ferredoxin in the physiological reaction. Furthemore, they decreased the k(cat)/K(m) value for NADPH in the ferricyanide reduction. Rapid-reaction (stopped-flow) kinetic analysis of the isolated reductive half-reaction of wild-type and His57Gln forms of FprA with NADPH and NADH allowed a detailed description of the mechanism of enzyme-bound FAD reduction, with the identification of the intermediates involved. The His57Gln mutation caused a 6-fold decrease in the rate of hydride transfer from either NADPH or NADH to the enzyme-bound FAD cofactor. The 3D structure of FprA-H57Q, obtained at 1.8 A resolution, explains the inefficient hydride transfer of the mutant in terms of a suboptimal geometry of the nicotinamide-isoalloxazine interaction in the active site. These data demonstrate the role of His57 in the correct binding of NADPH to FprA for the subsequent steps of the catalytic cycle to proceed at a high rate.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
20878669 H.Komori, D.Seo, T.Sakurai, and Y.Higuchi (2010).
Crystal structure analysis of Bacillus subtilis ferredoxin-NADP(+) oxidoreductase and the structural basis for its substrate selectivity.
  Protein Sci, 19, 2279-2290.
PDB codes: 3lzw 3lzx
19635450 H.Ouellet, J.B.Johnston, and P.R.Ortiz de Montellano (2010).
The Mycobacterium tuberculosis cytochrome P450 system.
  Arch Biochem Biophys, 493, 82-95.  
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.  
17635583 M.de 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.  
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 codes are shown on the right.

 

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