Molecular features of the L-type amino acid transporter 2 determine different import and export profiles for thyroid hormones and amino acids

Abstract

The L-type amino acid transporter 2 (LAT2) imports amino acids (AA) and also certain thyroid hormones (TH), e.g. 3,3'-T₂ and T₃, but not rT₃ and T₄. We utilized LAT2 mutations (Y130A, N133S, F242W) that increase 3,3'-T₂ import and focus here on import and export capacity for AA, T₄, T₃, BCH and derivatives thereof to delineate molecular features. Transport studies and analysis of competitive inhibition of import by radiolabelled TH and AA were performed in Xenopus laevis oocytes. Only Y130A, a pocket widening mutation, enabled import for T₄ and increased it for T₃. Mutant F242W showed increased 3,3'-T₂ import but no import rates for other TH derivatives. No export was detected for any TH by LAT2-wild type (WT). Mutations Y130A and N133S enabled only the export of 3,3'-T₂, while N133S also increased AA export. Thus, distinct molecular LAT2-features determine bidirectional AA transport but only an unidirectional 3,3'-T₂ and T₃ import.

Keywords: Competitive inhibition studies; Homology model; Molecular import and export; Site-directed mutagenesis; Thyroid hormone transport; Xenopus laevis oocytes.