The mechanisms responsible for L-tryptophan transport at both the maternal- and fetal-facing surfaces of the term placenta have been determined in isolated membrane vesicles as part of a study on placental indoleamine 2,3-dioxygenase, the L-tryptophan-catabolising enzyme recently shown to regulate feto-maternal immunology. Brush border vesicle uptake of L-tryptophan is substantially into an osmotically active space. It is sodium independent and N-ethylmaleimide sensitive. Uptake of L-tryptophan, which is markedly stereospecific, has a Km of 26.3 microM and Vmax of 1.72 pmol (mg protein)(-1) s(-1) and is completely abolished by the L-system-specific substrate 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH). These findings are in keeping with L-tryptophan transport being exclusively via system L (induced by the heterodimeric heavy chain of CD98 and system L-amino acid transporter-1 (LAT-1)). 1-Methyl-tryptophan (which is a known competitive inhibitor of indoleamine 2,3-dioxygenase) is a competitive inhibitor of L-tryptophan flux through this transport system (Ki = 113 microM). Basal membrane transport of L-tryptophan is more complex. Uptake is slower than at the brush border and although, as in the brush border, uptake is sodium independent, it is less sensitive to N-ethylmaleimide. There is clear evidence that two systems contribute to basal membrane transport since BCH is (in sodium-free media) only a partial inhibitor whereas L-histidine and L-cysteine are fully effective. The simplest explanation of these and other findings is that the basal membrane possesses two systems, one of which is similar to that induced by the heavy chain of CD98 and system L-amino acid transporter-2 (LAT-2). The other appears to be system y+L since in the presence of BCH inhibition by L-leucine but not by L-lysine is sodium dependent. These findings suggest the existence of non-identical carrier-mediated transport systems for L-tryptophan in brush border and basal membranes. This asymmetry may explain net transplacental transfer of this amino acid.