期刊
JOURNAL OF NEUROCHEMISTRY
卷 99, 期 1, 页码 20-28出版社
WILEY-BLACKWELL
DOI: 10.1111/j.1471-4159.2006.04003.x
关键词
coupling ions; neuronal glutamate transporter; radioactive uptake; reconstitution; substrate specificity; transport currents
资金
- NINDS NIH HHS [NS 16708] Funding Source: Medline
Glutamate transporters are essential for terminating synaptic transmission. Glutamate is translocated together with three sodium ions. In the neuronal glutamate transporter EAAC1, lithium can replace sodium. To address the question of whether the coupling ion interacts with the 'driven' substrate during co-transport, the kinetic parameters of transport of the three substrates, L-glutamate and D- and L-aspartate by EAAC-1 in sodium- and lithium-containing media were compared. The major effect of the substitution of sodium by lithium was on K-m. In the presence of sodium, the values for K-m and I-max of these substrates were similar. In the presence of lithium, the K-m for L-aspartate was increased around 13-fold. Remarkably, the corresponding increase for L-glutamate and D-aspartate was much larger, around 130-fold. In marked contrast, the K-i values for a non-transportable substrate analogue were similar in the presence of either sodium or lithium. The preference for L-aspartate in the presence of lithium was also observed when electrogenic transport of radioactive substrates was monitored in EAAC1-containing proteoliposomes. Our results indicate that, subsequent to substrate binding, the co-transported solutes interact functionally in the binding pocket of the transporter.
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