Identification, functional characterization and expression of a LAT type amino acid transporter from the mosquito Aedes aegypti.
We isolated two cDNAs from the mosquito Aedes aegypti, an L-amino acid transporter (AeaLAT) and a CD98 heavy chain (AeaCD98hc). Expression of AeaCD98hc or AeaLAT alone in Xenopus oocyte did not induce amino acid transport activity. However, co-expression of AeaCD98hc and AeaLAT, which are postulated to form a heterodimer protein linked through a disulfide bond, showed significant increase in amino acid transport activity. This heterodimeric protein showed uptake specificity for large neutral and basic amino acids. Small acidic neutral amino acids were poor substrates for this transporter. Neutral amino acid (leucine) uptake activity was partially Na+ dependent, because leucine uptake was approximately 44% lower in the absence of Na+ than in its presence. However, basic amino acid (lysine) uptake activity was completely Na+ independent at pH of 7.4. Extracellular amino acid concentration could be the main factor that determined amino acid transport. These results suggest the heteromeric protein is likely a uniporter mediating diffusion of amino acids in the absence of ions. The AeaLAT showed high level expression in the gastric caeca, Malpighian tubules and hindgut of larvae. In caeca and hindgut expression was in the apical cell membrane. However, in Malpighian tubules and in midgut, the latter showing low level expression, the transporter was detected in the basolateral membrane. This expression profile supports the conclusion that this AeaLAT is a nutrient amino acid transporter.
Jin
Xiangyang
X
Graduate Program in Environmental Toxicology, University of California, Riverside, CA 92521, USA.
Aimanova
Karlygash
K
Ross
Linda S
LS
Gill
Sarjeet S
SS
D000330:Aedes / Q000235:genetics* / Q000502:physiology*; D000595:Amino Acid Sequence; D027062:Amino Acid Transport System L / Q000096:biosynthesis* / Q000235:genetics*; D000596:Amino Acids / Q000493:pharmacokinetics; D000818:Animals; D017931:DNA Primers; D018076:DNA, Complementary; D004058:Diffusion; D004064:Digestive System / Q000737:chemistry; D004068:Digestive System Physiological Phenomena; D005786:Gene Expression Regulation; D019476:Insect Proteins / Q000096:biosynthesis* / Q000235:genetics*; D007814:Larva / Q000737:chemistry / Q000254:growth & development; D008969:Molecular Sequence Data; D009865:Oocytes; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D014981:Xenopus
D027062:Amino Acid Transport System L; D000596:Amino Acids; D017931:DNA Primers; D018076:DNA, Complementary; D019476:Insect Proteins