Characterization of Exosomal SLC22A5 (OCTN2) carnitine transporter

Scientific Reports
Lara ConsoleCesare Indiveri

Abstract

Exosomes are extracellular vesicles involved in cell-to-cell communication. Previous large scale proteomics revealed that they contain SLC proteins. However, no data on the function of exosomal SLCs is available, so far. An SLC localized in exosomes was here characterized for the first time: the carnitine transporter OCTN2 (SLC22A5). The protein was detected by Western Blot analysis in HEK293 exosomes. To investigate the functional properties of the exosomal OCTN2, the proteins extracted from vesicles were reconstituted into proteolipsomes and the transport function was measured as uptake of 3H-carnitine. Transport was stimulated by sodium and was dependent on pH. 3H-carnitine uptake was inhibited by Acetyl-carnitine, but not by Asn, Gln and Arg thus excluding interference by ATB0,+, an amino acid transporter which also recognizes carnitine. Cardiolipin failed to stimulate transport, excluding the activity of the mitochondrial Carnitine/acylcarnitine transporter. Increased level of exosomal OCTN2 was induced by treatment of HEK293 with the pro-inflammatory cytokine INFγ. All data concurred to demonstrate that OCTN2 present in exosomes is fully functional and is in its native conformation. Functional OCTN2 was detected also in h...Continue Reading

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Citations

Dec 28, 2019·The Journal of Pharmacology and Experimental Therapeutics·A David Rodrigues, Andrew Rowland
Sep 11, 2019·International Journal of Molecular Sciences·Lara ConsoleCesare Indiveri
Dec 20, 2019·Pharmacological Reviews·Hermann Koepsell
Oct 20, 2020·Frontiers in Cell and Developmental Biology·Lara ConsoleCesare Indiveri
Oct 10, 2020·Annual Review of Pharmacology and Toxicology·Adam S DarwichAmin Rostami-Hodjegan
Jan 30, 2020·Acta Pharmaceutica Sinica. B·Wenxin SongLigong Chen
Nov 1, 2021·Genome Biology and Evolution·Laura L ColbranJohn A Capra

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Methods Mentioned

BETA
glycosylation
assemble
electrophoresis

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