Publications

• Deorphaning a solute carrier 22 family member, SLC22A15, through functional genomic studies.

  Yee S.W., Buitrago D., Stecula A., Ngo H.X., Chien H.C., Zou L., Koleske M.L., Giacomini K.M.

  The human solute carrier 22A (SLC22A) family consists of 23 members, representing one of the largest families in the human SLC superfamily. Despite their pharmacological and physiological importance in the absorption and disposition of a range of solutes, eight SLC22A family members remain classif ... >> More

  The human solute carrier 22A (SLC22A) family consists of 23 members, representing one of the largest families in the human SLC superfamily. Despite their pharmacological and physiological importance in the absorption and disposition of a range of solutes, eight SLC22A family members remain classified as orphans. In this study, we used a multifaceted approach to identify ligands of orphan SLC22A15. Ligands of SLC22A15 were proposed based on phylogenetic analysis and comparative modeling. The putative ligands were then confirmed by metabolomic screening and uptake assays in SLC22A15 transfected HEK293 cells. Metabolomic studies and transporter assays revealed that SLC22A15 prefers zwitterionic compounds over cations and anions. We identified eight zwitterions, including ergothioneine, carnitine, carnosine, gabapentin, as well as four cations, including MPP⁺ , thiamine, and cimetidine, as substrates of SLC22A15. Carnosine was a specific substrate of SLC22A15 among the transporters in the SLC22A family. SLC22A15 transport of several substrates was sodium-dependent and exhibited a higher Km for ergothioneine, carnitine, and carnosine compared to previously identified transporters for these ligands. This is the first study to characterize the function of SLC22A15. Our studies demonstrate that SLC22A15 may play an important role in determining the systemic and tissue levels of ergothioneine, carnosine, and other zwitterions. << Less

  FASEB J. 34:15734-15752(2020) [PubMed] [EuropePMC]

  This publication is cited by 11 other entries.

• Functional characterization of monocarboxylate transporter 12 (SLC16A12/MCT12) as a facilitative creatine transporter.

  Takahashi M., Kishimoto H., Shirasaka Y., Inoue K.

  SLC16A12/MCT12 has been recently identified as a creatine transporter in a Xenopus oocyte expression system; however, the mechanism, by which MCT12 transports creatine, remains unclear. This study was performed to determine the functional and molecular characteristics of MCT12 in mammalian cells. ... >> More

  SLC16A12/MCT12 has been recently identified as a creatine transporter in a Xenopus oocyte expression system; however, the mechanism, by which MCT12 transports creatine, remains unclear. This study was performed to determine the functional and molecular characteristics of MCT12 in mammalian cells. The results showed that the uptake of [¹⁴C]creatine was not significantly increased in HEK293 cells transiently expressing MCT12 with or without CD147, a molecular chaperone, compared with mock cells. When [¹⁴C]creatine was accumulated in the cells with the aid of SLC6A8/CRT1, a concentrative creatine transporter, followed by assessing the remaining intracellular [¹⁴C]creatine after initiating efflux, coexpression of MCT12 resulted in a decrease in the intracellular [¹⁴C]creatine and remarkably enhanced the efflux of [¹⁴C]creatine from the cells in a time-dependent manner. This activity was not affected by extracellular pH. The creatine efflux activity involved dissipation by the mutations of conserved charged amino acids such as Arg37, Asp65 and Asp299 in the transmembrane domains, indicating direct involvement of MCT12 in the creatine efflux. These results suggest that MCT12 mediates facilitative diffusion of creatine, depending on the concentration gradient across the plasma membrane in mammalian cells. The finding may provide important clues to understanding the disposition kinetics of creatine and its derivatives. << Less

  Drug Metab. Pharmacokinet. 35:281-287(2020) [PubMed] [EuropePMC]

• Monocarboxylate transporter 12 as a guanidinoacetate efflux transporter in renal proximal tubular epithelial cells.

  Jomura R., Tanno Y., Akanuma S.I., Kubo Y., Tachikawa M., Hosoya K.I.

  Guanidinoacetate (GAA), which is a precursor of creatine, is mainly biosynthesized in the renal proximal tubular epithelial cells (RPTECs). Plasma concentration of GAA has been reported to be reduced in patients with monocarboxylate transporter 12 (MCT12) mutation (p.Q215X). However, the mechanism ... >> More

  Guanidinoacetate (GAA), which is a precursor of creatine, is mainly biosynthesized in the renal proximal tubular epithelial cells (RPTECs). Plasma concentration of GAA has been reported to be reduced in patients with monocarboxylate transporter 12 (MCT12) mutation (p.Q215X). However, the mechanism underlying GAA release from the RPTECs remains unclear. Therefore, to elucidate the role of MCT12 in renal GAA release, MCT12-mediated GAA transport was evaluated using the human and rat MCT12-expressing Xenopus laevis oocytes and primary-cultured rat RPTECs. [¹⁴C]GAA uptake by the human and rat MCT12-expressing oocytes was significantly higher than that by the water-injected oocytes. Rat MCT12-mediated uptake of [¹⁴C]GAA by the oocytes was found to be sodium ion (Na⁺)-independent and exhibited saturable kinetics with a Michaelis-Menten constant of 3.38 mM. Transport activities of rat MCT12 tend to increase along with increasing of extracellular pH. In addition, the efflux transport of [¹⁴C]GAA from the human and rat MCT12-expressing oocytes was significantly higher than that from the water-injected oocytes. These results suggest that both the influx and efflux transport of GAA is mediated by MCT12. In the primary-cultured rat RPTECs, [¹⁴C]GAA efflux transport was significantly reduced by the transfection of MCT12-specific siRNAs, suggesting that MCT12 participates in GAA efflux transport in rat RPTECs. Therefore, it suggests that MCT12 is involved in GAA release from RPTECs to the circulating blood, since MCT12 is known to be localized on the basal membrane of RPTECs. << Less

  Biochim. Biophys. Acta 1862:183434-183434(2020) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• The cataract and glucosuria associated monocarboxylate transporter MCT12 is a new creatine transporter.

  Abplanalp J., Laczko E., Philp N.J., Neidhardt J., Zuercher J., Braun P., Schorderet D.F., Munier F.L., Verrey F., Berger W., Camargo S.M., Kloeckener-Gruissem B.

  Creatine transport has been assigned to creatine transporter 1 (CRT1), encoded by mental retardation associated SLC6A8. Here, we identified a second creatine transporter (CRT2) known as monocarboxylate transporter 12 (MCT12), encoded by the cataract and glucosuria associated gene SLC16A12. A non-s ... >> More

  Creatine transport has been assigned to creatine transporter 1 (CRT1), encoded by mental retardation associated SLC6A8. Here, we identified a second creatine transporter (CRT2) known as monocarboxylate transporter 12 (MCT12), encoded by the cataract and glucosuria associated gene SLC16A12. A non-synonymous alteration in MCT12 (p.G407S) found in a patient with age-related cataract (ARC) leads to a significant reduction of creatine transport. Furthermore, Slc16a12 knockout (KO) rats have elevated creatine levels in urine. Transport activity and expression characteristics of the two creatine transporters are distinct. CRT2 (MCT12)-mediated uptake of creatine was not sensitive to sodium and chloride ions or creatine biosynthesis precursors, breakdown product creatinine or creatine phosphate. Increasing pH correlated with increased creatine uptake. Michaelis-Menten kinetics yielded a Vmax of 838.8 pmol/h/oocyte and a Km of 567.4 µm. Relative expression in various human tissues supports the distinct mutation-associated phenotypes of the two transporters. SLC6A8 was predominantly found in brain, heart and muscle, while SLC16A12 was more abundant in kidney and retina. In the lens, the two transcripts were found at comparable levels. We discuss the distinct, but possibly synergistic functions of the two creatine transporters. Our findings infer potential preventive power of creatine supplementation against the most prominent age-related vision impaired condition. << Less

  Hum. Mol. Genet. 22:3218-3226(2013) [PubMed] [EuropePMC]