PDBsum entry 1cc6

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protein ligands links
Oxidoreductase PDB id
Protein chain
391 a.a. *
Waters ×218
* Residue conservation analysis
PDB id:
Name: Oxidoreductase
Title: Phe161 and arg166 variants of p-hydroxybenzoate hydroxylase. Implications for NADPH recognition and structural stability
Structure: Protein (p-hydroxybenzoate hydroxylase). Chain: a. Engineered: yes. Mutation: yes
Source: Pseudomonas fluorescens. Organism_taxid: 294. Strain: tg2. Gene: poba. Expressed in: escherichia coli. Expression_system_taxid: 562.
Biol. unit: Dimer (from PQS)
2.20Å     R-factor:   0.185    
Authors: M.H.M.Eppink,C.Bunthof,H.A.Schreuder,W.J.H.Van Berkel
Key ref:
M.H.Eppink et al. (1999). Phe161 and Arg166 variants of p-hydroxybenzoate hydroxylase. Implications for NADPH recognition and structural stability. FEBS Lett, 443, 251-255. PubMed id: 10025942 DOI: 10.1016/S0014-5793(98)01726-8
04-Mar-99     Release date:   12-Mar-99    
Go to PROCHECK summary

Protein chain
Pfam   ArchSchema ?
P00438  (PHHY_PSEFL) -  p-hydroxybenzoate hydroxylase
394 a.a.
391 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 2 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: E.C.  - 4-hydroxybenzoate 3-monooxygenase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]

Benzoate Metabolism
      Reaction: 4-hydroxybenzoate + NADPH + O2 = protocatechuate + NADP+ + H2O
Bound ligand (Het Group name = PHB)
corresponds exactly
+ O(2)
= protocatechuate
+ NADP(+)
+ H(2)O
      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     metabolic process   5 terms 
  Biochemical function     oxidoreductase activity     4 terms  


DOI no: 10.1016/S0014-5793(98)01726-8 FEBS Lett 443:251-255 (1999)
PubMed id: 10025942  
Phe161 and Arg166 variants of p-hydroxybenzoate hydroxylase. Implications for NADPH recognition and structural stability.
M.H.Eppink, C.Bunthol, H.A.Schreuder, W.J.van Berkel.
Phe161 and Arg166 of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens belong to a newly discovered sequence motif in flavoprotein hydroxylases with a putative dual function in FAD and NADPH binding [1]. To study their role in more detail, Phe161 and Arg166 were selectively changed by site-directed mutagenesis. F161A and F161G are catalytically competent enzymes having a rather poor affinity for NADPH. The catalytic properties of R166K are similar to those of the native enzyme. R166S and R166E show impaired NADPH binding and R166E has lost the ability to bind FAD. The crystal structure of substrate complexed F161A at 2.2 A is indistinguishable from the native enzyme, except for small changes at the site of mutation. The crystal structure of substrate complexed R166S at 2.0 A revealed that Arg166 is important for providing an intimate contact between the FAD binding domain and a long excursion of the substrate binding domain. It is proposed that this interaction is essential for structural stability and for the recognition of the pyrophosphate moiety of NADPH.
  Selected figure(s)  
Figure 1.
Fig. 1. Stereoview of the Cα backbone of the enzyme-substrate complex of PHBH. 4-Hydroxybenzoate (POHB), cofactor (FAD), Phe^161 and Arg^166 are indicated.
Figure 2.
Fig. 2. Crystal structure of F161A in complex with POHB. A: Stereo diagram of the omit map of F161A, contoured at 3Ï, with the residues 160â164 omitted from the map. The atomic model of F161A is drawn in bold lines, while Phe^161 of wild-type PHBH is indicated in open lines. B: Superposition of the structures of wild-type PHBH and F161A in complex with POHB. The structure of the wild-type enzyme is drawn in open bonds and the structure of F161A is shown in solid bonds.
  The above figures are reprinted by permission from the Federation of European Biochemical Societies: FEBS Lett (1999, 443, 251-255) copyright 1999.  
  Figures were selected by the author.  

Literature references that cite this PDB file's key reference Google scholar

  PubMed id Reference
20055497 U.E.Ukaegbu, A.Kantz, M.Beaton, G.T.Gassner, and A.C.Rosenzweig (2010).
Structure and ligand binding properties of the epoxidase component of styrene monooxygenase .
  Biochemistry, 49, 1678-1688.
PDB code: 3ihm
11805318 J.Wang, M.Ortiz-Maldonado, B.Entsch, V.Massey, D.Ballou, and D.L.Gatti (2002).
Protein and ligand dynamics in 4-hydroxybenzoate hydroxylase.
  Proc Natl Acad Sci U S A, 99, 608-613.
PDB codes: 1k0i 1k0j 1k0l
11514508 D.Baitsch, C.Sandu, R.Brandsch, and G.L.Igloi (2001).
Gene cluster on pAO1 of Arthrobacter nicotinovorans involved in degradation of the plant alkaloid nicotine: cloning, purification, and characterization of 2,6-dihydroxypyridine 3-hydroxylase.
  J Bacteriol, 183, 5262-5267.  
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