p-Hydroxybenzoate hydroxylase from Pseudomonas fluorescens. 2. Fitting of the amino-acid sequence to the tertiary structure.
The complete primary and tertiary structure of p-hydroxybenzoate hydroxylase is now known. The amino acid sequences of the two largest CNBr peptides have been fitted to the electron-density map at 0.25-nm resolution. The parts of the polypeptide chain contributing the residues to the FAD-binding site and the residues of the substrate-binding site have been identified. The active site is located in a large hydrophobic area enclosed by all domains of the enzyme structure. Here the substrate, p-hydroxybenzoate, is bound near, but not in direct contact with, the isoalloxazine ring system of FAD. Many side chains from the C-terminal part of the polypeptide chain are involved in subunit-subunit interactions. In the center of one of the largely hydrophobic contact areas between the subunits, a cluster of six aromatic amino acids was found.
D015109:4-Hydroxybenzoate-3-Monooxygenase / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D001665:Binding Sites; D004583:Electrons; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000302:isolation & purification*; D008958:Models, Molecular; D011487:Protein Conformation; D011551:Pseudomonas fluorescens / Q000201:enzymology*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D005182:Flavin-Adenine Dinucleotide; D006899:Mixed Function Oxygenases; D015109:4-Hydroxybenzoate-3-Monooxygenase