Publications

• Choline transporter-like proteins 1 and 2 are newly identified plasma membrane and mitochondrial ethanolamine transporters.

  Taylor A., Grapentine S., Ichhpuniani J., Bakovic M.

  The membrane phospholipids phosphatidylcholine and phosphatidylethanolamine (PE) are synthesized de novo by the CDP-choline and CDP-ethanolamine (Kennedy) pathway, in which the extracellular substrates choline and ethanolamine are transported into the cell, phosphorylated, and coupled with diacylg ... >> More

  The membrane phospholipids phosphatidylcholine and phosphatidylethanolamine (PE) are synthesized de novo by the CDP-choline and CDP-ethanolamine (Kennedy) pathway, in which the extracellular substrates choline and ethanolamine are transported into the cell, phosphorylated, and coupled with diacylglycerol to form the final phospholipid product. Although multiple transport systems have been established for choline, ethanolamine transport is poorly characterized and there is no single protein assigned a transport function for ethanolamine. The solute carriers 44A (SLC44A) known as choline transporter-like proteins-1 and -2 (CTL1 and CTL2) are choline transporter at the plasma membrane and mitochondria. We report a novel function of CTL1 and CTL2 in ethanolamine transport. Using the lack or the gain of gene function in combination with specific antibodies and transport inhibitors we established two distinct ethanolamine transport systems of a high affinity, mediated by CTL1, and of a low affinity, mediated by CTL2. Both transporters are Na⁺-independent ethanolamine/H⁺ antiporters. Primary human fibroblasts with separate frameshift mutations in the CTL1 gene (M1= SLC44A1^ΔAsp517 and M2= SLC44A1^ΔSer126) are devoid of CTL1 ethanolamine transport but maintain unaffected CTL2 transport. The lack of CTL1 in M2 cells reduced the ethanolamine transport, the flux through the CDP-ethanolamine Kennedy pathway, and PE synthesis. In contrast, overexpression of CTL1 in M2 cells improved ethanolamine transport and PE synthesis. These data firmly establish that CTL1 and CTL2 are the first identified ethanolamine transporters in whole cells and mitochondria, with intrinsic roles in de novo PE synthesis by the Kennedy pathway and intracellular redistribution of ethanolamine. << Less

  J. Biol. Chem. 296:100604-100604(2021) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• The solute carrier 44A1 is a mitochondrial protein and mediates choline transport.

  Michel V., Bakovic M.

  Choline oxidation to betaine takes place in the mitochondria; however, a protein regulating mitochondrial choline transport was never identified. The purpose of this study was to analyze subcellular localization of the solute carrier 44A1 (SLC44A1), a plasma membrane choline transporter sensitive ... >> More

  Choline oxidation to betaine takes place in the mitochondria; however, a protein regulating mitochondrial choline transport was never identified. The purpose of this study was to analyze subcellular localization of the solute carrier 44A1 (SLC44A1), a plasma membrane choline transporter sensitive to inhibition by hemicholinium-3. We generated N- and C-terminal-SLC44A1-specific antibodies and analyzed localization of endogenous and overexpressed SLC44A1 in C2C12 mouse muscle cells, MCF7 human breast cancer cells, and mouse tissues using confocal microscopy, differential centrifugation, and Western blotting. We further performed choline uptake competition studies on isolated mitochondria using the specific inhibitor hemicholinium-3 and SLC44A1 antibodies, and analyzed mitochondria of FL83B hepatocytes after the targeted knock-down of SLC44A1 using siRNA technology. In addition, we analyzed SLC44A1 expression during choline deficiency. Localization studies revealed plasma membrane, cytosolic, microsomal, and mitochondrial localization of endogenous and His-tagged SLC44A1. Uptake studies in isolated mitochondria show an accumulation of (3)H-choline, which is strongly inhibited by hemicholinium-3 (60%), by an excess of unlabeled choline (97%), and by both SLC44A1 antibodies. SLC44A1 mRNA and protein expression were down-regulated during choline deficiency. These data clearly establish SLC44A1 as an important mediator of choline transport across both the plasma membrane and the mitochondrial membrane. << Less

  FASEB J. 23:2749-2758(2009) [PubMed] [EuropePMC]

• Molecular and functional characterization of choline transporter in rat renal tubule epithelial NRK-52E cells.

  Yabuki M., Inazu M., Yamada T., Tajima H., Matsumiya T.

  Homeostatic regulation of the plasma choline concentration depends on the effective functioning of a choline transporter in the kidney. However, the nature of the choline transport system in the kidney is poorly understood. In this study, we examined the molecular and functional characterization o ... >> More

  Homeostatic regulation of the plasma choline concentration depends on the effective functioning of a choline transporter in the kidney. However, the nature of the choline transport system in the kidney is poorly understood. In this study, we examined the molecular and functional characterization of choline uptake in the rat renal tubule epithelial cell line NRK-52E. Choline uptake was saturable and mediated by a single transport system, with an apparent Michaelis-Menten constant (K(m)) of 16.5 microM and a maximal velocity (V(max)) of 133.9 pmol/mg protein/min. The V(max) value of choline uptake was strongly enhanced in the absence of Na(+) without any change in K(m) values. The increase in choline uptake under Na(+)-free conditions was inhibited by Na(+)/H(+) exchanger (NHE) inhibitors. Choline uptake was inhibited by the choline uptake inhibitor hemicholinium-3 (HC-3) and organic cations, and was decreased by acidification of the extracellular medium and by intracellular alkalinization. Collapse of the plasma membrane H(+) electrochemical gradient by a protonophore inhibited choline uptake. NRK-52E cells mainly express mRNA for choline transporter-like proteins (CTL1 and CTL2), and NHE1 and NHE8. CTL1 protein was recognized in both plasma membrane and mitochondria. CTL2 protein was mainly expressed in mitochondria. The biochemical and pharmacological data indicated that CTL1 is functionally expressed in NRK-52E cells and is responsible for choline uptake. This choline transport system uses a directed H(+) gradient as a driving force, and its transport functions in co-operation with NHE8. Furthermore, the presence of CTL2 in mitochondria provides a potential site for the control of choline oxidation. << Less

  Arch. Biochem. Biophys. 485:88-96(2009) [PubMed] [EuropePMC]