PDBsum entry 3fwg

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protein ligands metals Protein-protein interface(s) links
Oxidoreductase PDB id
Protein chains
404 a.a. *
HEM ×2
CAM ×2
__K ×3
Waters ×949
* Residue conservation analysis
PDB id:
Name: Oxidoreductase
Title: Ferric camphor bound cytochrome p450cam, arg365leu, glu366gln, monoclinic crystal form
Structure: Camphor 5-monooxygenase. Chain: a, b. Synonym: cytochrome p450-cam, p450cam. Engineered: yes. Mutation: yes
Source: Pseudomonas putida. Organism_taxid: 303. Gene: camc, cyp101. Expressed in: escherichia coli. Expression_system_taxid: 562
1.55Å     R-factor:   0.186     R-free:   0.217
Authors: I.Schlichting,K.Von Koenig,C.Aldag,D.Hilvert
Key ref:
C.Aldag et al. (2009). Probing the role of the proximal heme ligand in cytochrome P450cam by recombinant incorporation of selenocysteine. Proc Natl Acad Sci U S A, 106, 5481-5486. PubMed id: 19293375 DOI: 10.1073/pnas.0810503106
18-Jan-09     Release date:   03-Mar-09    
Go to PROCHECK summary

Protein chains
Pfam   ArchSchema ?
P00183  (CPXA_PSEPU) -  Camphor 5-monooxygenase
415 a.a.
404 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 3 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: E.C.  - Camphor 5-monooxygenase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: +-camphor + reduced putidaredoxin + O2 = +-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
Bound ligand (Het Group name = CAM)
corresponds exactly
+ reduced putidaredoxin
+ O(2)
= (+)-exo-5-hydroxycamphor
+ oxidized putidaredoxin
+ H(2)O
      Cofactor: Heme-thiolate
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     cytoplasm   1 term 
  Biological process     oxidation-reduction process   2 terms 
  Biochemical function     oxidoreductase activity     7 terms  


DOI no: 10.1073/pnas.0810503106 Proc Natl Acad Sci U S A 106:5481-5486 (2009)
PubMed id: 19293375  
Probing the role of the proximal heme ligand in cytochrome P450cam by recombinant incorporation of selenocysteine.
C.Aldag, I.A.Gromov, I.García-Rubio, K.von Koenig, I.Schlichting, B.Jaun, D.Hilvert.
The unique monooxygenase activity of cytochrome P450cam has been attributed to coordination of a cysteine thiolate to the heme cofactor. To investigate this interaction, we replaced cysteine with the more electron-donating selenocysteine. Good yields of the selenoenzyme were obtained by bacterial expression of an engineered gene containing the requisite UGA codon for selenocysteine and a simplified yet functional selenocysteine insertion sequence (SECIS). The sulfur-to-selenium substitution subtly modulates the structural, electronic, and catalytic properties of the enzyme. Catalytic activity decreases only 2-fold, whereas substrate oxidation becomes partially uncoupled from electron transfer, implying a more complex role for the axial ligand than generally assumed.
  Selected figure(s)  
Figure 1.
The reaction path of P450cam. Compound I, a ferryl-oxo π-cation porphyrin radical, is the putative oxidant that reacts directly with substrate. Dashed arrows indicate possible uncoupling processes.
Figure 3.
Structure of C357U P450cam* in the camphor-bound state. (A) Active site of the selenoenzyme (yellow) superimposed on that of P450cam* (green) and WT P450cam (1dz4, gray); all 3 structures were determined from the monoclinic crystal form. Although the axial cysteine ligand at position 357 was substituted by selenocysteine, the structural integrity of the enzyme is maintained. The increased Fe-Se distance may account for the 2 side chain conformations observed for Leu-358 in the selenoenzyme. The camphor-binding site is not affected by the mutations (see Fig. S2). The camphor molecule has been omitted for clarity. (B) View of the region around Gln-366 in C357U P450cam* (yellow), P450cam* (green), and the WT protein (gray). Mutating the native glutamate residue to glutamine induces displacement of the connected chain of ordered water molecules.
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
21350756 J.A.Gámez, and M.Yáñez (2011).
Is Se-Se bond cleavage the most favourable process in electron attachment to diselenides? The importance of asymmetry.
  Chem Commun (Camb), 47, 3939-3941.  
20446763 T.C.Pochapsky, S.Kazanis, and M.Dang (2010).
Conformational plasticity and structure/function relationships in cytochromes P450.
  Antioxid Redox Signal, 13, 1273-1296.  
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