7US3

Structure of Putrescine N-hydroxylase Involved Complexed with NADP+

7US3 の概要

• エントリーDOI: 10.2210/pdb7us3/pdb
• 分子名称: Putrescine N-hydroxylase, FLAVIN-ADENINE DINUCLEOTIDE, NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE, ... (6 entities in total)
• 機能のキーワード: monooxygenase, putrescine n-hydroxylase, flavoprotein
• 由来する生物種: Acinetobacter baumannii
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 220844.13

構造登録者

Tanner, J.J.,Bogner, A.N. (登録日: 2022-04-22, 公開日: 2022-11-30, 最終更新日: 2023-10-25)

主引用文献

Lyons, N.S.,Bogner, A.N.,Tanner, J.J.,Sobrado, P.
Kinetic and Structural Characterization of a Flavin-Dependent Putrescine N -Hydroxylase from Acinetobacter baumannii.
Biochemistry, 61:2607-2620, 2022

PubMed Abstract: is a Gram-negative opportunistic pathogen that causes nosocomial infections, especially among immunocompromised individuals. The rise of multidrug resistant strains of has limited the use of standard antibiotics, highlighting a need for new drugs that exploit novel mechanisms of pathogenicity. Disrupting iron acquisition by inhibiting the biosynthesis of iron-chelating molecules (siderophores) secreted by the pathogen is a potential strategy for developing new antibiotics. Here we investigated FbsI, an -hydroxylating monooxygenase involved in the biosynthesis of fimsbactin A, the major siderophore produced by FbsI was characterized using steady-state and transient-state kinetics, spectroscopy, X-ray crystallography, and small-angle X-ray scattering. FbsI was found to catalyze the -hydroxylation of the aliphatic diamines putrescine and cadaverine. Maximum coupling of the reductive and oxidative half-reactions occurs with putrescine, suggesting it is the preferred () substrate. FbsI uses both NADPH and NADH as the reducing cofactor with a slight preference for NADPH. The crystal structure of FbsI complexed with NADP was determined at 2.2 Å resolution. The structure exhibits the protein fold characteristic of Class B flavin-dependent monooxygenases. FbsI is most similar in 3D structure to the cadaverine -hydroxylases DesB and DfoA. Small-angle X-ray scattering shows that FbsI forms a tetramer in solution like the -hydroxylating monooxygenases of the SidA/IucD/PvdA family. A model of putrescine docked into the active site provides insight into substrate recognition. A mechanism for the catalytic cycle is proposed where dehydration of the C4a-hydroxyflavin intermediate is partially rate-limiting, and the hydroxylated putrescine product is released before NADP.

PubMed: 36314559
DOI: 10.1021/acs.biochem.2c00493

実験手法

X-RAY DIFFRACTION (2.2 Å)