Identification and characterization of 3-iodothyronamine intracellular transport.
3-Iodothyronamine (T(1)AM) is a naturally occurring thyroid hormone metabolite with distinct biological effects that are opposite those of thyroid hormone. The known molecular targets of T(1)AM include both plasma membrane and intracellular proteins, suggesting that intracellular transport of T(1)AM may be an important component of its action, although no uptake mechanism has yet been described. Using various human cell lines, we show that, indeed, cellular uptake of T(1)AM occurs in multiple cell types and that this process involves specific, saturable, and inhibitable transport mechanisms. These mechanisms are sodium and chloride independent, pH dependent, thyronamine specific, and do not involve the likely candidate transporters of other monoamines, organic cations, or thyroid hormones. A large-scale RNA interference screen targeting the entire solute carrier superfamily of transporter genes reveals that the transport of T(1)AM into cells involves multiple transporters, and we identify eight transporters that may contribute to the uptake of T(1)AM in HeLa cells. This type of transporter small interfering RNA screening approach can be used in general to identify the constellation of transporters that participate in the intracellular disposition of compounds.
Ianculescu
Alexandra G
AG
Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, California 94143, USA.
Giacomini
Kathleen M
KM
Scanlan
Thomas S
TS
D015655:1-Methyl-4-phenylpyridinium / Q000378:metabolism; D000212:Acyclovir / Q000378:metabolism; D001692:Biological Transport / Q000235:genetics / Q000502:physiology*; D002331:Carnitine / Q000378:metabolism; D002460:Cell Line; D033722:Equilibrative-Nucleoside Transporter 2 / Q000235:genetics / Q000378:metabolism; D004970:Estrone / Q000031:analogs & derivatives / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism; D027501:Monocarboxylic Acid Transporters / Q000235:genetics / Q000378:metabolism; D027362:Organic Anion Transport Protein 1 / Q000235:genetics / Q000378:metabolism; D029362:Organic Anion Transporters, Sodium-Independent / Q000235:genetics / Q000378:metabolism; D027702:Organic Cation Transporter 1 / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000502:physiology; D027981:Symporters; D019789:Tetraethylammonium / Q000378:metabolism; D013970:Thyronines / Q000378:metabolism*
C487948:3-iodothyronamine; D033722:Equilibrative-Nucleoside Transporter 2; D026901:Membrane Transport Proteins; D027501:Monocarboxylic Acid Transporters; D027362:Organic Anion Transport Protein 1; D029362:Organic Anion Transporters, Sodium-Independent; D027702:Organic Cation Transporter 1; D034741:RNA, Small Interfering; C090735:SLC16A2 protein, human; D027981:Symporters; D013970:Thyronines; C114338:organic anion transport protein 3; D004970:Estrone; C004547:thyronamine; D019789:Tetraethylammonium; C017296:estrone sulfate; D015655:1-Methyl-4-phenylpyridinium; D002331:Carnitine; D000212:Acyclovir