Two-component carnitine monooxygenase from Escherichia coli: functional characterization, inhibition and mutagenesis of the molecular interface.
Carnitine monooxygenase
Rieske-type oxygenase
gut microbiota
l-carnitine
site-directed mutagenesis
trimethylamine
Journal
Bioscience reports
ISSN: 1573-4935
Titre abrégé: Biosci Rep
Pays: England
ID NLM: 8102797
Informations de publication
Date de publication:
30 09 2022
30 09 2022
Historique:
received:
18
05
2022
revised:
16
08
2022
accepted:
06
09
2022
pubmed:
7
9
2022
medline:
24
9
2022
entrez:
6
9
2022
Statut:
ppublish
Résumé
Gut microbial production of trimethylamine (TMA) from l-carnitine is directly linked to cardiovascular disease. TMA formation is facilitated by carnitine monooxygenase, which was proposed as a target for the development of new cardioprotective compounds. Therefore, the molecular understanding of the two-component Rieske-type enzyme from Escherichia coli was intended. The redox cofactors of the reductase YeaX (FMN, plant-type [2Fe-2S] cluster) and of the oxygenase YeaW (Rieske-type [2Fe-2S] and mononuclear [Fe] center) were identified. Compounds meldonium and the garlic-derived molecule allicin were recently shown to suppress microbiota-dependent TMA formation. Based on two independent carnitine monooxygenase activity assays, enzyme inhibition by meldonium or allicin was demonstrated. Subsequently, the molecular interplay of the reductase YeaX and the oxygenase YeaW was addressed. Chimeric carnitine monooxygenase activity was efficiently reconstituted by combining YeaX (or YeaW) with the orthologous oxygenase CntA (or reductase CntB) from Acinetobacter baumannii. Partial conservation of the reductase/oxygenase docking interface was concluded. A structure guided mutagenesis approach was used to further investigate the interaction and electron transfer between YeaX and YeaW. Based on AlphaFold structure predictions, a total of 28 site-directed variants of YeaX and YeaW were kinetically analyzed. Functional relevance of YeaX residues Arg271, Lys313 and Asp320 was concluded. Concerning YeaW, a docking surface centered around residues Arg83, Lys104 and Lys117 was hypothesized. The presented results might contribute to the development of TMA-lowering strategies that could reduce the risk for cardiovascular disease.
Identifiants
pubmed: 36066069
pii: 231753
doi: 10.1042/BSR20221102
pmc: PMC9508527
pii:
doi:
Substances chimiques
Disulfides
0
Methylamines
0
Methylhydrazines
0
Sulfinic Acids
0
allicin
3C39BY17Y6
3-(2,2,2-trimethylhydrazine)propionate
73H7UDN6EC
Flavin Mononucleotide
7N464URE7E
Mixed Function Oxygenases
EC 1.-
Oxidoreductases
EC 1.-
Oxygenases
EC 1.13.-
trimethylamine
LHH7G8O305
Carnitine
S7UI8SM58A
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
© 2022 The Author(s).
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