Identification of a novel conserved sequence motif in flavoprotein hydroxylases with a putative dual function in FAD/NAD(P)H binding

• Published in: Protein science 1997-11, Vol.6 (11), p.2454-2458
• Main Authors: Eppink, Michel H. M., Berkel, Willem J. H. Van, Schreuder, Herman A.
• Format: Article
• Language: English
• Subjects: 4-Hydroxybenzoate-3-Monooxygenase - chemistry; Amino Acid Sequence; Binding Sites; Conserved Sequence; fingerprint; Flavin-Adenine Dinucleotide; flavoprotein family; Flavoproteins - chemistry; Mixed Function Oxygenases - chemistry; Models, Molecular; Molecular Sequence Data; NAD; NADP; NADPH‐binding; p‐hydroxybenzoate hydroxylase; Sequence Alignment; Sequence Homology, Amino Acid
• ISSN: 0961-8368; 1469-896X
• DOI: 10.1002/pro.5560061119

A novel conserved sequence motif has been located among the flavoprotein hydroxylases. Based on the crystal structure and site‐directed mutagenesis studies of p‐hydroxybenzoate hydroxylase (PHBH) from Pseudomonas fluorescens, this amino acid fingerprint sequence is proposed to play a dual function in both FAD and NAD(P)H binding. In PHBH, the novel sequence motif (residues 153‐166) includes strand A4 and the N‐terminal part of helix H7. The conserved amino acids Asp 159, Gly 160, and Arg 166 are necessary for maintaining the structure. The backbone oxygen of Cys 158 and backbone nitrogens of Gly 160 and Phe 161 interact indirectly with the pyrophosphate moiety of FAD, whereas it is known from mutagenesis studies that the side chain of the moderately conserved His 162 is involved in NADPH binding.