Characterization of 3-[125I]iodo-alpha-methyl-L-tyrosine transport via human L-type amino acid transporter 1.
We examined transport of 3-[(125)I]iodo-alpha-methyl-L-tyrosine ([(125)I]IMT) in Xenopus laevis oocytes co-expressing human L-type amino acid transporter 1 (a component of system L) and human 4F2hc. Human LAT1 mediated transport of [(125)I]IMT. [(125)I]IMT uptake was decreased by the presence of L-isomers of Cys, Leu, Ileu, Phe, Met, Tyr, His, Trp and Val and D-isomers of Leu, Phe and Met. Human LAT1-mediated [(125)I]IMT uptake was highly stereoselective for the L-isomers of Tyr, His, Trp, Val and Ileu. To examine the effects of 3-iodination and alpha-methylation on IMT transport, kinetic parameters of IMT were compared with those of mother Tyr and 3-[(125)I]iodo-L-tyrosine (3-I-Tyr). Uptake of Tyr, 3-I-Tyr and [(125)I]IMT followed Michaelis-Menten kinetics, with K(m) values of 29.0 +/- 5.1, 12.6 +/- 6.1 and 22.6 +/- 4.1 microM, respectively. Neither the alpha-methyl group nor the size of the 3-iodinated Tyr residue was an obstacle to transport via hLAT1. Furthermore, affinity of IMT for hLAT1 is higher than that of the natural parent tyrosine. The level of efflux mediated by hLAT1 was highly stimulated by extracellularly applied L-Leu, suggesting exchange of [(125)I]IMT and L-Leu via hLAT1.
Shikano
Naoto
N
Department of Radiological Sciences, Ibaraki Prefectural University of Health Sciences, 4669-2 Ami Ami-machi, Inashiki-gun, Ibaraki 300-0394, Japan. sikano@ipu.ac.jp
Kanai
Yoshikatsu
Y
Kawai
Keiichi
K
Ishikawa
Nobuyoshi
N
Endou
Hitoshi
H
D027062:Amino Acid Transport System L / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D005260:Female; D027282:Fusion Regulatory Protein 1, Heavy Chain / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007536:Isomerism; D008781:Methyltyrosines / Q000737:chemistry / Q000493:pharmacokinetics*; D009865:Oocytes / Q000737:chemistry / Q000000981:diagnostic imaging* / Q000378:metabolism*; D011877:Radionuclide Imaging; D019275:Radiopharmaceuticals / Q000737:chemistry / Q000493:pharmacokinetics; D011994:Recombinant Proteins / Q000378:metabolism; D012016:Reference Values; D014443:Tyrosine / Q000737:chemistry; D014982:Xenopus laevis
D027062:Amino Acid Transport System L; D027282:Fusion Regulatory Protein 1, Heavy Chain; D008781:Methyltyrosines; D019275:Radiopharmaceuticals; D011994:Recombinant Proteins; D014443:Tyrosine; C030855:3-iodo-alpha-methyltyrosine