Molecular mechanism of inhibition of the mitochondrial carnitine/acylcarnitine transporter by omeprazole revealed by proteoliposome assay, mutagenesis and bioinformatics

PloS One
Annamaria TonazziCesare Indiveri

Abstract

The effect of omeprazole on the mitochondrial carnitine/acylcarnitine transporter has been studied in proteoliposomes. Externally added omeprazole inhibited the carnitine/carnitine antiport catalysed by the transporter. The inhibition was partially reversed by DTE indicating that it was caused by the covalent reaction of omeprazole with Cys residue(s). Inhibition of the C-less mutant transporter indicated also the occurrence of an alternative non-covalent mechanism. The IC50 of the inhibition of the WT and the C-less CACT by omeprazole were 5.4 µM and 29 µM, respectively. Inhibition kinetics showed non competitive inhibition of the WT and competitive inhibition of the C-less. The presence of carnitine or acylcarnitines during the incubation of the proteoliposomes with omeprazole increased the inhibition. Using site-directed Cys mutants it was demonstrated that C283 and C136 were essential for covalent inhibition. Molecular docking of omeprazole with CACT indicated the formation of both covalent interactions with C136 and C283 and non-covalent interactions in agreement with the experimental data.

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Citations

Oct 16, 2015·Biochimica Et Biophysica Acta·Nicola GiangregorioCesare Indiveri
Apr 8, 2015·Chemical Research in Toxicology·Annamaria TonazziCesare Indiveri
Dec 11, 2014·Toxicological Sciences : an Official Journal of the Society of Toxicology·Michele GalluccioCesare Indiveri
Feb 20, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Nicola GiangregorioCesare Indiveri
Sep 19, 2020·PloS One·Naw May Pearl Cartee, Michael M Wang
Jun 30, 2019·SLAS Discovery·Mariafrancesca ScaliseCesare Indiveri
Jul 25, 2019·Biochimica Et Biophysica Acta. Bioenergetics·Nicola GiangregorioCesare Indiveri

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Methods Mentioned

BETA
PCR

Software Mentioned

Deep View
LowModeMD
MOE Conformational Search
Molecular Operating Environment ( MOE )
MOE Site Finder
CACT
MOE Builder
Quantity One
MOE
SwissModel

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