Cystine/glutamate transporter
Cystine/glutamate transporter is an antiporter that in humans is encoded by the SLC7A11 gene.
The SLC7A11 gene encodes a sodium-independent cystine-glutamate antiporter that is chloride dependent, also known as xCT. Along with a heavy chain subunit from SLC3A2, the SLC7A11 light chain comprises system Xc-, which is the functional cystine-glutamate antiporter. While the SLC3A2 heavy chain is a chaperone for many other light chains that participate in amino acid transport, the SLC7A11 light chain is specific for system Xc-, and the terms xCT/SLC7A11 and system Xc- are used interchangeably in much of the literature.
SLC7A11 plays an important role in glutathione production throughout nervous and non-nervous tissues. In the nervous system, SLC7A11 regulates synaptic activity by stimulating extrasynaptic receptors and performs nonvesicular glutamate release. This gene is highly expressed by astrocytes and couples the uptake of one molecule of cystine with the release of one molecule of glutamate. The expression of Xc- was detected throughout the brain with higher expression found in the basolateral amygdala, the retina and the prefrontal cortex. The inhibition of system Xc- has been found to alter a number of behaviors, which suggests that it plays a key role in excitatory signaling.
Structure
SLC7A11 is a member of a heterodimeric Na+-independent anionic amino acid transport system highly specific for cystine and glutamate. This antiporter imports cystine and exports glutamate, which are both amino acids. An antiporter functions with a one-to-one counter-transport, which is when one substance is transported across the membrane at the same time another substance is transported across the membrane in the opposite direction. The antiporter is a heterodimeric amino acid transporter, requiring both the SLC7A11 light chain and SLC3A2 heavy chain linked by a disulfide bridge. The SLC7A11 light chain has 12 transmembrane domains consisting of 501 amino acids, and the SLC3A2 heavy chain appears to be highly conserved among transporters. The human SLC7A11 has an 89% similarity of amino acids to the homologous mouse xCT protein. The complementary DNA, cDNA, has a total of 9648 base pairs. The SLC7A11 gene has been found not only in the brain, but has also been found to be expressed in the spinal cord, pancreas, and in glioma cells.Regulation
There are many mechanisms that exist to regulate the expression of system Xc-, although it is not the sole determinant of extracellular glutamate or intracellular glutathione. An example is amino acid deprivation, which triggers up regulation of the transporter. A key regulator is extracellular glutamate; when it becomes excessive, it goes from an excitatory transmitter to an excitotoxin. The inhibition of uptake of extracellular cystine into cells leads to decreased levels of intracellular glutathione which leads to ferroptosis. This regulation may be done through Excitatory Amino Acid Transporters, which decrease extracellular glutamate and increase intracellular glutamate in astrocytes. When looking at its structure, xCT seems to be the main determinant for the system's activity. Glutamate and cystine can be transported in both directions, but, generally, more cystine is imported and more glutamate is exported. Extracellular glutamate acts as a competitive inhibitor for cystine uptake via system Xc-.Glutamate
There is a high amount of glutamate in mammalian cells. Glutamate is necessary for excitatory signaling between neurons. The release must be highly organized, due to the large amounts of glutamate at the synaptic cleft, and the fact that it is released at high speeds. This mechanism of release at the synaptic cleft is partially controlled through the active transport of glutamate out of astrocytes by system Xc-. This release also has a physiological role in the regulation of glutamatergic metabotropic receptors and control of other neurotransmitters.It has been demonstrated that, in the embryonic retina, Xc- exchanger is responsible for 50% of total glutamate uptake, representing a Sodium-Independent system within this tissue. The high activity of Xc- in the retina is correlated to a neuroprotective role, once it can take up excessive extracellular glutamate and provide precursors for the synthesis of Glutathione.
Cystine
Cystine is a dimer consisting of two cysteine molecules and the formation of a disulfide bond. This amino acid is a rate limiting substrate used in the SLC7A11 cystine/glutamate transporter and is usually imported into the cell. Cysteine-158 is specifically used in the formation of the disulfide bridge for the protein structure of system Xc-. There are neurotoxins, such as BMAA, that can prevent the intake of cystine, which can lead to decreased extracellular glutamate levels and an increase in oxidative stress.Pharmacological Inhibition
System Xc- can be inhibited by many small molecules. Excess amounts of the endogenous substrate glutamate inhibits the function of system Xc-. Synthetic small molecules such as erastin, sulfasalazine, and sorafenib can inhibit system Xc- function and induce ferroptosis.Clinical relevancy
Many central nervous system disorders are due to a dysfunction in glutamate signaling. Glutamate is transported via EAATs and system Xc-. If either of these transporters are impaired, it could result in a disruption in glutamate homeostasis and lead to a variety of CNS disordersDrug addiction
It has been found that cocaine produces a decrease in Cystine-Glutamate exchange via system Xc-, leading to a decrease in basal, extra synaptic glutamate levels in the nucleus accumbens core region of the brains of cocaine-withdrawn rats. It has also been observed in withdrawn rats that a decrease in Group 2 mGluR inhibition of vesicular release, most likely due to the decrease in extrasynaptic glutamate levels, leads to an increase in cocaine-evoked glutamate signaling in their NAcc. An infusion of cysteine in the NAcc of withdrawn rats leads to an increase in extrasynaptic glutamate, near the levels of the control rats, and prevents an increase in synaptic glutamate signaling after a cocaine injection. These findings suggest there is a decrease in system Xc- activity in cocaine-withdrawn rats. It has also been found that cocaine increases glutamate signaling in the synaptic cleft, further supporting this conclusion.Administration of the cysteine prodrugs N-acetylcysteine or L-2-oxothiazolidine-4-carboxylate blocks cocaine reinstatement in rats. N-acetylcysteine has been shown to decrease drug-seeking behavior for nicotine and heroin as well. However, N-acetylcysteine does not alter the cocaine-induced rush or euphoria; it only causes a reduction in drug-seeking behavior. N-acetylcysteine works by increasing levels of cysteine in cells, leading to an increase in system Xc- activity. This increase in system Xc- activity leads to an increase in extrasynaptic glutamate, causing stimulation of Group 2 mGluRs and an inhibition of synaptic release of glutamate. Cysteine prodrugs also lead to an increase in antioxidant properties by increasing levels of glutathione. Increased levels of glutathione lead to a lower toxicity of methamphetamine and alcohol, and cause a decrease in tumor formation after chronic smoking. N-acetylcysteine has been shown to decrease cravings and use of cocaine and tobacco, as well as other compulsive behaviors such as gambling and trichotillomania.
Repeated administration of cocaine causes disruptions in glutamate homeostasis that lead to a decrease in function of EAATs. It is also possible that glutamate is diffusing from surrounding synapses and is stimulating extrasynaptic receptors. All of these factors may be leading to the disruptions in glutamate signaling that are associated with drug addiction.
Schizophrenia
It has been proposed that schizophrenia may be due to an increase or a decrease in glutamate signaling, leading to abnormal excitatory signaling in the prefrontal cortex region of the brain. Glutamate release by astrocytes has been linked to the synchrony of neurons in the hippocampus and cortex. A decrease in system Xc- activity may result in an increase in synaptic glutamate and a decrease in extrasynaptic glutamate. Administration of N-acetylcysteine leads to an increase in extrasynaptic NMDA receptor activation, suggesting that glutamate released from system Xc- may cause the activation of extrasynaptic NMDA receptors. A decrease in system Xc- activity may cause a decrease in the activation of extrasynaptic NMDA receptors due to either a decrease in extrasynaptic Glutamate levels or a decrease in glutathione levels after the decrease in cystine transport. On the other hand, a decrease in system Xc- activity may lead to an increase in the activation of synaptic NMDA receptors due to the decrease in activation of Group 2 mGluRs. A decrease in nonvesicular release of glutamate leads to an increase in expression of postsynaptic glutamate receptors, such as NMDA receptors. A disruption in nonvesicular glutamate release may affect synapse formation, lead to altered release of neurotransmitters, and could even disturb cortical migration during development. All of these seem to be associated with schizophrenia.An increase in the expression of Group 2 mGluRs, which could arise from a chronic under stimulation of these receptors, has been associated with schizophrenia. An increase in levels of system Xc- has also been found in postmortem schizophrenia patients, indicating that there may have been a decrease in net function of these receptors as well, leading to greater expression. It has been observed that Schizophrenia patients have a decreased level of glutathione in their prefrontal cortex, further supporting the conclusion that system Xc- may not be functioning properly.
Clinical trials have shown therapeutic potential for N-acetylcysteine in treating schizophrenia. Again, changes in EAATs due to disruptions in Glutamate homeostasis may also be involved.
Recent study showed that mRNA expression levels of both SLC3A2 and SLC7A11 in WBCs of schizophrenia patients are lower than that of healthy individuals. The finding supports the hypo-glutamatergic neurotransmission hypothesis in schizophrenia.