Caveolin-1--a novel interacting partner of organic cation/carnitine transporter (Octn2): effect of protein kinase C on this interaction in rat astrocytes

PloS One
Magdalena CzeredysKatarzyna A Nałecz

Abstract

OCTN2--the Organic Cation Transporter Novel family member 2 (SLC22A5) is known to be a xenobiotic/drug transporter. It transports as well carnitine--a compound necessary for oxidation of fatty acids and mutations of its gene cause primary carnitine deficiency. Octn2 regulation by protein kinase C (PKC) was studied in rat astrocytes--cells in which β-oxidation takes place in the brain. Activation of PKC with phorbol ester stimulated L-carnitine transport and increased cell surface presence of the transporter, although no PKC-specific phosphorylation of Octn2 could be detected. PKC activation resulted in an augmented Octn2 presence in cholesterol/sphingolipid-rich microdomains of plasma membrane (rafts) and increased co-precipitation of Octn2 with raft-proteins, caveolin-1 and flotillin-1. Deletion of potential caveolin-1 binding motifs pointed to amino acids 14-22 and 447-454 as the caveolin-1 binding sites within Octn2 sequence. A direct interaction of Octn2 with caveolin-1 in astrocytes upon PKC activation was detected by proximity ligation assay, while such an interaction was excluded in case of flotillin-1. Functioning of a multi-protein complex regulated by PKC has been postulated in rOctn2 trafficking to the cell surface, ...Continue Reading

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Citations

May 16, 2014·Archives of Biochemistry and Biophysics·Katarzyna MichalecKatarzyna A Nałęcz
Apr 5, 2017·International Journal of Molecular Sciences·Abdullah MayatiOlivier Fardel
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Jul 13, 2020·Journal of Neurochemistry·Barbara JuraszekKatarzyna A Nałęcz
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Aug 10, 2016·Neurochemical Research·Katarzyna A Nałęcz
Jun 3, 2020·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Yuchen ZhangBruno Hagenbuch
Nov 17, 2020·Frontiers in Cell and Developmental Biology·Katarzyna A Nałęcz

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Methods Mentioned

BETA
reverse transcription PCR
PCR
transfection
PMA
flow cytometry
electrophoresis
immunoprecipitation
proximity ligation assay
transmission electron microscopy
proximity ligation

Software Mentioned

GraphPad Prism
GraphPad

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