PDBsum entry 1d7l

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Oxidoreductase PDB id
Protein chain
394 a.a. *
SO4 ×3
Waters ×349
* Residue conservation analysis
PDB id:
Name: Oxidoreductase
Title: Structure-function correlations of the reaction of reduced nicotinamide analogs with p-hydroxybenzoate hydroxylase sub with a series of 8-substituted flavins
Structure: P-hydroxybenzoate hydroxylase. Chain: a. Engineered: yes
Source: Pseudomonas aeruginosa. Organism_taxid: 287. Expressed in: escherichia coli. Expression_system_taxid: 562.
Biol. unit: Dimer (from PQS)
2.20Å     R-factor:   0.184     R-free:   0.235
Authors: M.Ortiz-Maldonado,D.Gatti,D.P.Ballou,V.Massey
Key ref:
M.Ortiz-Maldonado et al. (1999). Structure-function correlations of the reaction of reduced nicotinamide analogues with p-hydroxybenzoate hydroxylase substituted with a series of 8-substituted flavins. Biochemistry, 38, 16636-16647. PubMed id: 10600126 DOI: 10.1021/bi991603u
18-Oct-99     Release date:   21-Jan-00    
Go to PROCHECK summary

Protein chain
Pfam   ArchSchema ?
P20586  (PHHY_PSEAE) -  p-hydroxybenzoate hydroxylase
394 a.a.
394 a.a.
Key:    PfamA domain  Secondary structure  CATH domain

 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 = RFL) matches with 92.86% similarity
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: 10.1021/bi991603u Biochemistry 38:16636-16647 (1999)
PubMed id: 10600126  
Structure-function correlations of the reaction of reduced nicotinamide analogues with p-hydroxybenzoate hydroxylase substituted with a series of 8-substituted flavins.
M.Ortiz-Maldonado, D.Gatti, D.P.Ballou, V.Massey.
Structural and kinetic studies have revealed two flavin conformations in p-hydroxybenzoate hydroxylase (PHBH), the in-position and the out-position. Conversion between these two conformations is believed to be essential during catalysis. Although substrate hydroxylation occurs while the flavin in PHBH is in the in-conformation, the position of the flavin during reduction by NADPH is uncertain. To investigate the catalytic importance of the out-conformation of the flavin and to clarify the mechanism of flavin reduction in PHBH, we report quantitative structure-reactivity relationships (QSAR) using PHBH substituted separately with nine derivatives of FAD modified in the 8-position and four dihydronicotinamide analogues as reducing agents. The 8-position of the FAD isoalloxazine ring was chosen for modification because in PHBH it has minimal interactions with the protein and is accessible to solvent. The chemical sequence of events during catalysis by PHBH was not altered when using any of the modified flavins, and normal products were obtained. Although the rate of reduction of PHBH reconstituted with flavin derivatives is expected to be dependent on the redox potential of the flavin, no strict correlation was observed. Instead, the rate of reduction correlated with the kappa-substituent constant, which is based on size and hydrophobicity of the 8-substituent on the FAD. Substituents that sterically hinder attainment of the out-conformation decreased the rate of flavin reduction much more than expected on the basis of the redox potential of the flavin. The results of this QSAR analysis are consistent with the hypothesis that the flavin in PHBH must move to the out-conformation for proper formation of the charge-transfer complex between NADPH and FAD that is necessary for rapid flavin reduction.

Literature references that cite this PDB file's key reference

  PubMed id Reference
20450491 B.Calamini, K.Ratia, M.G.Malkowski, M.Cuendet, J.M.Pezzuto, B.D.Santarsiero, and A.D.Mesecar (2010).
Pleiotropic mechanisms facilitated by resveratrol and its metabolites.
  Biochem J, 429, 273-282.  
17226839 Y.K.Choe, S.Nagase, and K.Nishimoto (2007).
Theoretical study of the electronic spectra of oxidized and reduced states of lumiflavin and its derivative.
  J Comput Chem, 28, 727-739.  
10819972 M.B.Murataliev, and R.Feyereisen (2000).
Interaction of NADP(H) with oxidized and reduced P450 reductase during catalysis. Studies with nucleotide analogues.
  Biochemistry, 39, 5066-5074.  
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