The D-amino acid transport by the invertebrate SLC6 transporters KAAT1 and CAATCH1 from Manduca sexta.

Abstract

The ability of the SLC6 family members, the insect neutral amino acid cotransporter KAAT1(K+‐coupled amino acid transporter 1) and its homologous CAATCH1(cation anion activated amino acid transporter/channel), to transport D‐amino acids has been investigated through heterologous expression in Xenopus laevis oocytes and electrophysiological techniques. In the presence of D‐isomers of leucine, serine, and proline, the ms KAAT1 generates inward, transport‐associated, currents with variable relative potencies, depending on the driving ion Na+ or K+. Higher concentrations of D‐leucine (≥1 mmol/L) give rise to an anomalous response that suggests the existence of a second binding site with inhibitory action on the transport process. ms CAATCH1 is also able to transport the D‐amino acids tested, including D‐leucine, whereas L‐leucine acts as a blocker. A similar behavior is exhibited by the KAAT1 mutant S308T, confirming the relevance of the residue in this position in L‐leucine binding and the different interaction of D‐leucine with residues involved in transport mechanism. D‐leucine and D‐serine on various vertebrate orthologs B0 AT1 (SLC6A19) elicited only a very small current and singular behavior was not observed, indicating that it is specific of the insect neutral amino acid transporters. These findings highlight the relevance of D‐amino acid absorption in the insect nutrition and metabolism and may provide new evidences in the molecular transport mechanism of SLC6 family.