Solute carrier family 1, member 1 (SLC1A1; also known as EAAT3 and EAAC1) is the major epithelial transporter of glutamate and aspartate in the kidneys and intestines of rodents. Within the brain, SLC1A1 serves as the predominant neuronal glutamate transporter and buffers the synaptic release of the excitatory neurotransmitter glutamate within the interneuronal synaptic cleft. Recent studies have also revealed that polymorphisms in SLC1A1 are associated with obsessive-compulsive disorder (OCD) in early-onset patient cohorts. Here we report that SLC1A1 mutations leading to substitution of arginine to tryptophan at position 445 (R445W) and deletion of isoleucine at position 395 (I395del) cause human dicarboxylic aminoaciduria, an autosomal recessive disorder of urinary glutamate and aspartate transport that can be associated with mental retardation. These mutations of conserved residues impeded or abrogated glutamate and cysteine transport by SLC1A1 and led to near-absent surface expression in a canine kidney cell line. These findings provide evidence that SLC1A1 is the major renal transporter of glutamate and aspartate in humans and implicate SLC1A1 in the pathogenesis of some neurological disorders.

Calcium transport was monitored by measuring ATP-dependent 45Ca uptake into membrane vesicles prepared from rabbit lens cortex. Calcium-stimulated adenosinetriphosphatase (Ca2(+)-ATPase) activity was also measured in the same membrane preparation. Both uptake and Ca2(+)-ATPase activity were inhibited by vanadate. Calcium activation of the uptake process was similar to that of the Ca2(+)-ATPase. Calcium uptake was prevented by calcium ionophore A23187, suggesting that the calcium transported into the vesicles remains diffusible. The ATP-dependent calcium uptake probably represents the transport of calcium into "inside-out" membrane vesicles by the Ca2(+)-ATPase mechanism that normally shifts calcium outward from the lens cytoplasm. The temperature dependence of the Ca2(+)-ATPase and the calcium uptake process was determined. Because lipid order can influence Ca2(+)-ATPase function, we attempted to correlate calcium transport with the physical state of the membrane lipids. Infrared spectroscopy was used to determine the temperature dependence of the CH2 symmetric stretching frequency (an order parameter) in the lipids. A similarity was noted between the temperature-dependence curves for lipid order, Ca2(+)-ATPase, and calcium uptake rate. Entropy, enthalpy, and transition temperature calculated for the Ca2(+)-ATPase and calcium uptake process were in the same range as those parameters calculated for the lipid-phase transition.

No abstract