8EEJ

C. ammoniagenes monoamine oxidase (MAO) C424S variant bound to dopamine

8EEJ の概要

• エントリーDOI: 10.2210/pdb8eej/pdb
• 分子名称: Amine oxidase, L-DOPAMINE, FLAVIN-ADENINE DINUCLEOTIDE, ... (4 entities in total)
• 機能のキーワード: flavoenzyme, substrate, flavoprotein
• 由来する生物種: Corynebacterium ammoniagenes
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 101890.28

構造登録者

Muellers, S.N.,Allen, K.N. (登録日: 2022-09-07, 公開日: 2023-02-01, 最終更新日: 2023-10-25)

主引用文献

Muellers, S.N.,Tararina, M.A.,Kuzmanovic, U.,Galagan, J.E.,Allen, K.N.
Structural Insights into the Substrate Range of a Bacterial Monoamine Oxidase.
Biochemistry, 62:851-862, 2023

PubMed Abstract: Monoamine oxidases (MAOs) play a key role in the breakdown of primary and secondary amines. In eukaryotic organisms, these enzymes are vital to the regulation of monoamine neurotransmitters and the degradation of dietary monoamines. MAOs have also been identified in prokaryotic species, although their role in these organisms is not well understood. Here, we report the biophysical and structural properties of a promiscuous, bacterial MAO from (MAO). MAO catalyzes the oxidation of a number of monoamine substrates including dopamine and norepinephrine, as well as exhibiting some activity with polyamine substrates such as cadaverine. The X-ray crystal structures of Michaelis complexes with seven substrates show that conserved hydrophobic interactions and hydrogen-bonding pattern (for polar substrates) allow the broad specificity range. The structure of MAO identifies an unusual cysteine (Cys424) residue in the so-called "aromatic cage", which flanks the flavin isoalloxazine ring in the active site. Site-directed mutagenesis, steady-state kinetics in air-saturated buffer, and UV-vis spectroscopy revealed that Cys424 plays a role in the pH dependence and modulation of electrostatics within the MAO active site. Notably, bioinformatic analysis shows a propensity for variation at this site within the "aromatic cage" of the flavin amine oxidase (FAO) superfamily. Structural analysis also identified the conservation of a secondary substrate inhibition site, present in a homologous member of the superfamily. Finally, genome neighborhood diagram analysis of MAO in the context of the FAO superfamily allows us to propose potential roles for these bacterial MAOs in monoamine and polyamine degradation and catabolic pathways related to scavenging of nitrogen.

PubMed: 36662673
DOI: 10.1021/acs.biochem.2c00540

実験手法

X-RAY DIFFRACTION (1.54 Å)