pobA

Summary

Gene Symbol: pobA
Description: 4-hydroxybenzoate 3-monooxygenase
Species: Pseudomonas aeruginosa PAO1

Top Publications

  1. ncbi 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
    Department of Biological Chemistry, University of Michigan, Ann Arbor 48109 0606, USA
    Biochemistry 38:16636-47. 1999
  2. ncbi Crystal structure of p-hydroxybenzoate hydroxylase
    R K Wierenga
    J Mol Biol 131:55-73. 1979
  3. ncbi Crystal structures of wild-type p-hydroxybenzoate hydroxylase complexed with 4-aminobenzoate,2,4-dihydroxybenzoate, and 2-hydroxy-4-aminobenzoate and of the Tyr222Ala mutant complexed with 2-hydroxy-4-aminobenzoate. Evidence for a proton channel and a new
    H A Schreuder
    BIOSON Research Institute, University of Groningen, The Netherlands
    Biochemistry 33:10161-70. 1994
  4. ncbi Structure and function of mutant Arg44Lys of 4-hydroxybenzoate hydroxylase implications for NADPH binding
    M H Eppink
    Department of Biochemistry, Agricultural University, Wageningen, The Netherlands
    Eur J Biochem 231:157-65. 1995
  5. pmc Crystal structure of p-hydroxybenzoate hydroxylase reconstituted with the modified FAD present in alcohol oxidase from methylotrophic yeasts: evidence for an arabinoflavin
    W J van Berkel
    Department of Biochemistry, Agricultural University, Wageningen, The Netherlands
    Protein Sci 3:2245-53. 1994
  6. ncbi The mobile flavin of 4-OH benzoate hydroxylase
    D L Gatti
    Department of Biological Chemistry, University of Michigan, Ann Arbor 48109
    Science 266:110-4. 1994
  7. ncbi Changes in the catalytic properties of p-hydroxybenzoate hydroxylase caused by the mutation Asn300Asp
    B A Palfey
    Department of Biological Chemistry, University of Michigan, Ann Arbor 48109 0606
    Biochemistry 33:1545-54. 1994
  8. ncbi Crystal structures of mutant Pseudomonas aeruginosa p-hydroxybenzoate hydroxylases: the Tyr201Phe, Tyr385Phe, and Asn300Asp variants
    M S Lah
    Department of Biological Chemistry, University of Michigan, Ann Arbor 48109
    Biochemistry 33:1555-64. 1994
  9. ncbi pH-dependent structural changes in the active site of p-hydroxybenzoate hydroxylase point to the importance of proton and water movements during catalysis
    D L Gatti
    Department of Biological Chemistry, University of Michigan, Ann Arbor 48109, USA
    Biochemistry 35:567-78. 1996
  10. ncbi Lys42 and Ser42 variants of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens reveal that Arg42 is essential for NADPH binding
    M H Eppink
    Department of Biomolecular Sciences, Wageningen Agricultural University, The Netherlands
    Eur J Biochem 253:194-201. 1998

Detail Information

Publications35

  1. ncbi 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
    Department of Biological Chemistry, University of Michigan, Ann Arbor 48109 0606, USA
    Biochemistry 38:16636-47. 1999
    ....
  2. ncbi Crystal structure of p-hydroxybenzoate hydroxylase
    R K Wierenga
    J Mol Biol 131:55-73. 1979
  3. ncbi Crystal structures of wild-type p-hydroxybenzoate hydroxylase complexed with 4-aminobenzoate,2,4-dihydroxybenzoate, and 2-hydroxy-4-aminobenzoate and of the Tyr222Ala mutant complexed with 2-hydroxy-4-aminobenzoate. Evidence for a proton channel and a new
    H A Schreuder
    BIOSON Research Institute, University of Groningen, The Netherlands
    Biochemistry 33:10161-70. 1994
    ..It is proposed that movement of the FAD out of the active site may provide an entrance for the substrate to enter the active site and an exit for the product to leave...
  4. ncbi Structure and function of mutant Arg44Lys of 4-hydroxybenzoate hydroxylase implications for NADPH binding
    M H Eppink
    Department of Biochemistry, Agricultural University, Wageningen, The Netherlands
    Eur J Biochem 231:157-65. 1995
    ..Replacement of Arg44 by Lys however affects NADPH binding, resulting in a low yield of the charge-transfer species between reduced flavin and NADP+. It is inferred from these data that Arg44 is indispensable for optimal catalysis...
  5. pmc Crystal structure of p-hydroxybenzoate hydroxylase reconstituted with the modified FAD present in alcohol oxidase from methylotrophic yeasts: evidence for an arabinoflavin
    W J van Berkel
    Department of Biochemistry, Agricultural University, Wageningen, The Netherlands
    Protein Sci 3:2245-53. 1994
    ..7 mol of hydrogen peroxide per mol NADPH oxidized. It is proposed that flavin motion in p-hydroxybenzoate hydroxylase is important for efficient reduction and that the flavin "out" conformation is associated with the oxidase activity...
  6. ncbi The mobile flavin of 4-OH benzoate hydroxylase
    D L Gatti
    Department of Biological Chemistry, University of Michigan, Ann Arbor 48109
    Science 266:110-4. 1994
    ..Movement of the flavin appears to be essential for the translocation of substrates and products into the solvent-shielded active site during catalysis...
  7. ncbi Changes in the catalytic properties of p-hydroxybenzoate hydroxylase caused by the mutation Asn300Asp
    B A Palfey
    Department of Biological Chemistry, University of Michigan, Ann Arbor 48109 0606
    Biochemistry 33:1545-54. 1994
    ....
  8. ncbi Crystal structures of mutant Pseudomonas aeruginosa p-hydroxybenzoate hydroxylases: the Tyr201Phe, Tyr385Phe, and Asn300Asp variants
    M S Lah
    Department of Biological Chemistry, University of Michigan, Ann Arbor 48109
    Biochemistry 33:1555-64. 1994
    ..The functional consequences of these changes in the enzyme structure and of the introduction of the carboxyl group at 300 are described and discussed in the accompanying paper (Palfey et al., 1994b)...
  9. ncbi pH-dependent structural changes in the active site of p-hydroxybenzoate hydroxylase point to the importance of proton and water movements during catalysis
    D L Gatti
    Department of Biological Chemistry, University of Michigan, Ann Arbor 48109, USA
    Biochemistry 35:567-78. 1996
    ..Transfer of water between the chain of proton donors and the solvent also appears to be an essential part of the mechanism that provides reversible transfer of protons during the hydroxylation reaction...
  10. ncbi Lys42 and Ser42 variants of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens reveal that Arg42 is essential for NADPH binding
    M H Eppink
    Department of Biomolecular Sciences, Wageningen Agricultural University, The Netherlands
    Eur J Biochem 253:194-201. 1998
    ..In spite of this, the Arg42 variants fully couple enzyme reduction to substrate hydroxylation. Sequence-comparison studies suggest that Arg42 is involved in binding of the 2'-phosphoadenosine moiety of NADPH...
  11. ncbi Interdomain binding of NADPH in p-hydroxybenzoate hydroxylase as suggested by kinetic, crystallographic and modeling studies of histidine 162 and arginine 269 variants
    M H Eppink
    Department of Biomolecular Sciences, Laboratory of Biochemistry, Wageningen Agricultural University, Dreijenlaan 3, 6703 HA Wageningen, The Netherlands
    J Biol Chem 273:21031-9. 1998
    ..An interdomain binding mode for NADPH is proposed which takes a novel sequence motif (Eppink, M. H. M., Schreuder, H. A., and van Berkel, W. J. H. (1997) Protein Sci. 6, 2454-2458) into account...
  12. ncbi Phe161 and Arg166 variants of p-hydroxybenzoate hydroxylase. Implications for NADPH recognition and structural stability
    M H Eppink
    Department of Biomolecular Sciences, Wageningen University Research Centre, The Netherlands
    FEBS Lett 443:251-5. 1999
    ..It is proposed that this interaction is essential for structural stability and for the recognition of the pyrophosphate moiety of NADPH...
  13. ncbi Switch of coenzyme specificity of p-hydroxybenzoate hydroxylase
    M H Eppink
    Department of Biomolecular Sciences, Laboratory of Biochemistry, Wageningen University, Wageningen, 6703 HA, The Netherlands
    J Mol Biol 292:87-96. 1999
    ..This is the first report on the coenzyme reversion of a flavoprotein aromatic hydroxylase...
  14. pmc Protein and ligand dynamics in 4-hydroxybenzoate hydroxylase
    Jian Wang
    Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201, USA
    Proc Natl Acad Sci U S A 99:608-13. 2002
    ..This work clearly shows how complex dynamics can play a central role in catalysis by enzymes...
  15. ncbi Oxygen reactions in p-hydroxybenzoate hydroxylase utilize the H-bond network during catalysis
    Mariliz Ortiz-Maldonado
    Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109 0606, USA
    Biochemistry 43:15246-57. 2004
    ..1, indicating the involvement of the H-bond network. We conclude that product deprotonation enhances the rate of a specific conformational change required for both product release and the elimination of water from C4a-OH-FAD...
  16. ncbi Removal of a methyl group causes global changes in p-hydroxybenzoate hydroxylase
    Lindsay J Cole
    Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109 0606, USA
    Biochemistry 44:8047-58. 2005
    ..This work demonstrates some general principles of how enzymes use conformational movements to allow both access and egress of substrates and product, while restricting access to the solvent at a critical stage in catalysis...
  17. ncbi Comparison of the three-dimensional protein and nucleotide structure of the FAD-binding domain of p-hydroxybenzoate hydroxylase with the FAD- as well as NADPH-binding domains of glutathione reductase
    R K Wierenga
    J Mol Biol 167:725-39. 1983
    ..Since they can stabilize a negative charge around O-2 alpha, they may be important for the catalytic processes...
  18. ncbi Kinetics of proton-linked flavin conformational changes in p-hydroxybenzoate hydroxylase
    Kendra King Frederick
    Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109 0606, USA
    Biochemistry 44:13304-14. 2005
    ..These results allow a detailed kinetic scheme to be proposed for the reductive half-reaction of the wild-type enzyme. Three kinetic models considered for substrate-induced isomerization are analyzed in the Appendix...
  19. ncbi The amino-acid sequence of the three smallest CNBr peptides from p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens
    J M Vereijken
    Eur J Biochem 113:151-7. 1980
    ..CB3, CB4 and CB5, were determined by automated Edman degradation and analysis of enzymatic subdigests. These peptides form a continuous stretch of 110 residues from the N terminus: (Formula: See Text)...
  20. ncbi Crystallization and preliminary x-ray investigation of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens
    J Drenth
    J Biol Chem 250:5268-9. 1975
    ..5, 39% saturated ammonium sulfate, and 1mM p-hydroxybenzoate. The space group is C2221 with 8 molecules/unit cell. When p-hydroxybenzoate is removed, the diffraction pattern and the cell dimensions change...
  21. ncbi Crystal structure of the reduced form of p-hydroxybenzoate hydroxylase refined at 2.3 A resolution
    H A Schreuder
    BIOSON Research Institute, University of Groningen, The Netherlands
    Proteins 14:178-90. 1992
    ..The position of the substrate is virtually unaltered with respect to its position in the oxidized enzyme. No trace of a bound NADP+ or NADPH molecule was found...
  22. ncbi Catalytic function of tyrosine residues in para-hydroxybenzoate hydroxylase as determined by the study of site-directed mutants
    B Entsch
    Department of Biochemistry, Microbiology, and Nutrition, Uniersity of New England, Armidale, New South Wales, Australia
    J Biol Chem 266:17341-9. 1991
    ..Tyr-385----Phe reacts with oxygen to form 25% oxidized enzyme, and 75% flavin hydroperoxide, which successfully hydroxylates the substrate. This mutant also hydroxylates the product (3, 4-dihydroxybenzoate) to form gallic acid...
  23. ncbi Engineering of microheterogeneity-resistant p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens
    K Eschrich
    Department of Biochemistry, Agricultural University, Wageningen, The Netherlands
    FEBS Lett 277:197-9. 1990
    ..Crystals of the C116S mutant are isomorphous with the crystal form of wild-type enzyme. A difference electron density confirms the mutation made...
  24. ncbi Analysis of the active site of the flavoprotein p-hydroxybenzoate hydroxylase and some ideas with respect to its reaction mechanism
    H A Schreuder
    Laboratory of Chemical Physics, Groningen, The Netherlands
    Biochemistry 29:3101-8. 1990
    ..Polarization of the peroxide oxygen-oxygen bond by the enzyme may enhance the reactivity of flavin 4a-peroxide...
  25. ncbi Sequence and organization of pobA, the gene coding for p-hydroxybenzoate hydroxylase, an inducible enzyme from Pseudomonas aeruginosa
    B Entsch
    Department of Biochemistry, Microbiology and Nutrition, University of New England, Armidale, N S W, Australia
    Gene 71:279-91. 1988
    The only recognized gene for the metabolism of p-hydroxybenzoate in Pseudomonads (pobA) has been isolated from Pseudomonas aeruginosa to provide the DNA for mutagenesis studies of the protein product, p-hydroxybenzoate hydroxylase...
  26. ncbi Crystal structure of the p-hydroxybenzoate hydroxylase-substrate complex refined at 1.9 A resolution. Analysis of the enzyme-substrate and enzyme-product complexes
    H A Schreuder
    Laboratory of Chemical Physics, University of Groningen, The Netherlands
    J Mol Biol 208:679-96. 1989
    ..Six out of eight peptide residues near the flavin ring are oriented with their nitrogen atom pointing towards the ring.(ABSTRACT TRUNCATED AT 400 WORDS)..
  27. ncbi The coenzyme analogue adenosine 5-diphosphoribose displaces FAD in the active site of p-hydroxybenzoate hydroxylase. An x-ray crystallographic investigation
    J M Van Der Laan
    Laboratory of Chemical Physics, University of Groningen, The Netherlands
    Biochemistry 28:7199-205. 1989
    ..1 M potassium phosphate buffer, pH 8.0. The empty pocket left by the flavin ring is filled by solvent, leaving the architecture of the active site and the binding of the substrate largely unaffected...
  28. ncbi The influence of purification and protein heterogeneity on the crystallization of p-hydroxybenzoate hydroxylase
    J M Van Der Laan
    Laboratory of Chemical Physics, University of Groningen, The Netherlands
    Eur J Biochem 179:715-24. 1989
    ..Sulfite and dithiothreitol improve crystallization by dissociating the enzyme oligomers into dimers; sulfite especially gives excellent results...
  29. ncbi Molecular modeling reveals the possible importance of a carbonyl oxygen binding pocket for the catalytic mechanism of p-hydroxybenzoate hydroxylase
    H A Schreuder
    Laboratory of Chemical Physics, Groningen, The Netherlands
    J Biol Chem 263:3131-6. 1988
    ..The possible consequences of this model for the reaction mechanism of p-hydroxybenzoate hydroxylase are discussed...
  30. ncbi Crystal structure of p-hydroxybenzoate hydroxylase complexed with its reaction product 3,4-dihydroxybenzoate
    H A Schreuder
    Laboratory of Chemical Physics, University of Groningen, The Netherlands
    J Mol Biol 199:637-48. 1988
    ....
  31. ncbi p-Hydroxybenzoate hydroxylase from Pseudomonas fluorescens. 2. Fitting of the amino-acid sequence to the tertiary structure
    W J Weijer
    Eur J Biochem 133:109-18. 1983
    ..In the center of one of the largely hydrophobic contact areas between the subunits, a cluster of six aromatic amino acids was found...
  32. ncbi p-Hydroxybenzoate hydroxylase from Pseudomonas fluorescens. 1. Completion of the elucidation of the primary structure
    J Hofsteenge
    Eur J Biochem 133:91-108. 1983
    ..An important tool in the analysis of the amino acid sequence of peptide CB1 was the proteinase Lys-C from Lysobacter enzymogenes, which preferentially cleaves at lysine residues...
  33. ncbi Primary and tertiary structure studies of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens. Isolation and alignment of the CNBr peptides; interactions of the protein with flavin adenine dinucleotide
    J Hofsteenge
    Eur J Biochem 113:141-50. 1980
    ..D-amino acid oxidase. This finding suggests the presence of a mononucleotide binding fold at the N terminus of the latter...
  34. ncbi Substitution of Arg214 at the substrate-binding site of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens
    W van Berkel
    Department of Biochemistry, Agricultural University, Wageningen, The Netherlands
    Eur J Biochem 210:411-9. 1992
    ..From spectral and kinetic results it is suggested that secondary binding of the substrate occurs at the re side of the flavin, where the nicotinamide moiety of NADPH is supposed to bind...
  35. ncbi Primary structure of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens
    W J Weijer
    Biochim Biophys Acta 704:385-8. 1982
    ..K., de Jong, R.J., Kalk, K.H., Hol, W.G.J. and Drenth, J. (1979) J. Mol. Biol. 131, 55-73) and from protein sequence analysis. The polypeptide chain of the monomer contains 394 amino acids and has a molecular weight of 44 299...