3WE0

L-Amino acid oxidase/monooxygenase from Pseudomonas sp. AIU 813

Summary for 3WE0

• Entry DOI: 10.2210/pdb3we0/pdb
• Descriptor: L-amino acid oxidase/monooxygenase, FLAVIN-ADENINE DINUCLEOTIDE (3 entities in total)
• Functional Keywords: flavin-containing monoamine oxidase family, rossmann fold, oxidoreductase (oxidase and monooxygenase), oxidoreductase
• Biological source: Pseudomonas sp.
• Total number of polymer chains: 2
• Total formula weight: 130604.84

Authors

Im, D.H.,Matsui, D.,Fukuta, Y.,Fushinobu, S.,Isobe, K.,Asano, Y. (deposition date: 2013-06-26, release date: 2014-02-12, Last modification date: 2024-03-20)

Primary citation

Matsui, D.,Im, D.H.,Sugawara, A.,Fukuta, Y.,Fushinobu, S.,Isobe, K.,Asano, Y.
Mutational and crystallographic analysis of l-amino acid oxidase/monooxygenase from Pseudomonas sp. AIU 813: Interconversion between oxidase and monooxygenase activities
FEBS Open Bio, 4:220-228, 2014

PubMed Abstract: In this study, it was shown for the first time that l-amino acid oxidase of Pseudomonas sp. AIU813, renamed as l-amino acid oxidase/monooxygenase (l-AAO/MOG), exhibits l-lysine 2-monooxygenase as well as oxidase activity. l-Lysine oxidase activity of l-AAO/MOG was increased in a p-chloromercuribenzoate (p-CMB) concentration-dependent manner to a final level that was five fold higher than that of the non-treated enzyme. In order to explain the effects of modification by the sulfhydryl reagent, saturation mutagenesis studies were carried out on five cysteine residues, and we succeeded in identifying l-AAO/MOG C254I mutant enzyme, which showed five-times higher specific activity of oxidase activity than that of wild type. The monooxygenase activity shown by the C254I variant was decreased significantly. Moreover, we also determined a high-resolution three-dimensional structure of l-AAO/MOG to provide a structural basis for its biochemical characteristics. The key residue for the activity conversion of l-AAO/MOG, Cys-254, is located near the aromatic cage (Trp-418, Phe-473, and Trp-516). Although the location of Cys-254 indicates that it is not directly involved in the substrate binding, the chemical modification by p-CMB or C254I mutation would have a significant impact on the substrate binding via the side chain of Trp-516. It is suggested that a slight difference of the binding position of a substrate can dictate the activity of this type of enzyme as oxidase or monooxygenase.

PubMed: 24693490
DOI: 10.1016/j.fob.2014.02.002

Experimental method

X-RAY DIFFRACTION (1.9 Å)