Lysine uptake by cloned hCAT-2B: comparison with hCAT-1 and with trophoblast surface membranes.
To study the cationic amino-acid transporter hCAT-2B of human placenta, total RNA was harvested from primary cultured trophoblast and from the BeWo choriocarcinoma cell line (b30 clone) and used for reverse transcription (RT) and polymerase chain reaction (PCR). Primers based on published sequences identified expression of mRNA for hCAT-2B. RT-PCR yielded a 2.06 kb hCAT-2B cDNA, which was cloned. hCAT-2B cRNA injection into Xenopus laevis oocytes stimulated saturable lysine uptake (Km approximately 125 mM). In the presence of Na+, uptake was completely inhibited by L-arginine but only partially by neutral amino acids. To compare directly the interaction of hCAT-1 and hCAT-2B with neutral amino acids and sodium, we examined the inhibition of these transporters by L-leucine and L-alanine over a wide concentration range. L-Alanine and L-leucine inhibit uptake by hCAT-2B substantially less completely than uptake by hCAT-1. The interaction of hCAT-2B resembles that of system y+ in the microvillous membrane of human placenta, while that of hCAT-1 is more comparable to that of system y+ in basal membrane. The identification and characterization of the various cationic amino-acid transporters of the human placenta have the potential to increase the understanding of the cellular mechanism of transplacental transfer.
Furesz
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Department of Pediatrics, Box 8116, Washington University School of Medicine at St. Louis Children's Hospital, One Children's Place, St. Louis, MO 63110, USA.
Heath-Monnig
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Kamath
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Smith
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D000409:Alanine / Q000494:pharmacology; D026923:Amino Acid Transport Systems, Basic; D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D027201:Cationic Amino Acid Transporter 1 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D027202:Cationic Amino Acid Transporter 2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002999:Clone Cells; D007930:Leucine / Q000494:pharmacology; D008239:Lysine / Q000493:pharmacokinetics*; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D015203:Reproducibility of Results; D014327:Trophoblasts / Q000187:drug effects / Q000378:metabolism*; D014982:Xenopus laevis / Q000235:genetics / Q000378:metabolism
D026923:Amino Acid Transport Systems, Basic; D000596:Amino Acids; D027201:Cationic Amino Acid Transporter 1; D027202:Cationic Amino Acid Transporter 2; C420825:SLC7A1 protein, human; C414177:SLC7A2 protein, human; D007930:Leucine; D008239:Lysine; D000409:Alanine