Ligand bound structure of a 6-hydroxynicotinic acid 3-monooxygenase provides mechanistic insights

被引：1

作者：

Turlington, Zachary R. ^{[1
]}

de Macedo, Sofia Vaz Ferreira ^{[1
]}

Perry, Kay ^{[2
,3
]}

Belsky, Sam L. ^{[4
]}

Faust, Jennifer A. ^{[4
]}

Snider, Mark J. ^{[4
]}

Hicks, Katherine A. ^{[1
]}

机构：

[1] SUNY Coll Cortland, Dept Chem, Cortland, NY 13045 USA

[2] NE CAT, Argonne, IL USA

[3] Cornell Univ, Dept Chem & Chem Biol, Argonne Natl Lab, Argonne, IL USA

[4] Coll Wooster, Dept Chem, Wooster, OH 44691 USA

来源：

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS | 2024年 / 752卷

基金：

美国国家卫生研究院;

关键词：

Flavin monooxygenase; Bacterial nicotinic acid metabolism; Flavin adenine dinucleotide (FAD); Protein structure; P-HYDROXYBENZOATE HYDROXYLASE; WASTE-WATER CONTAMINANTS; NATIONAL RECONNAISSANCE; CRYSTAL-STRUCTURE; UNITED-STATES; FLAVIN; PHARMACEUTICALS; DEGRADATION; SUBSTRATE; PROTEIN;

D O I：

10.1016/j.abb.2023.109859

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

6-Hydroxynicotinic acid 3-monooxygenase (NicC) is a bacterial enzyme involved in the degradation of nicotinic acid. This enzyme is a Class A flavin-dependent monooxygenase that catalyzes a unique decarboxylative hydroxylation. The unliganded structure of this enzyme has previously been reported and studied using steady- and transient-state kinetics to support a comprehensive kinetic mechanism. Here we report the crystal structure of the H47Q NicC variant in both a ligand-bound (solved to 2.17 angstrom resolution) and unliganded (1.51 angstrom resolution) form. Interestingly, in the liganded form, H47Q NicC is bound to 2-mercaptopyridine (2-MP), a contaminant present in the commercial stock of 6-mercaptopyridine-3-carboxylic acid(6-MNA), a substrate analogue. 2-MP binds weakly to H47Q NicC and is not a substrate for the enzyme. Based on kinetic and thermodynamic characterization, we have fortuitously captured a catalytically inactive H47Q NicC center dot 2-MP complex in our crystal structure. This complex reveals interesting mechanistic details about the reaction catalyzed by 6-hydroxynicotinic acid 3monooxygenase.

引用

页数：8

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