Photoreceptors transiently release glutamate in response to changing levels of light and dark. Excitatory amino acid transporters (EAATs) modulate glutamatergic signal transduction by removing glutamate from the synaptic cleft. In axon terminals of photoreceptors two functionally distinct EAAT isoforms co-localize, the retina specific EAAT5 and a splice variant of EAAT2, GLT1c. Whereas GLT1c is an effective glutamate transporter with small associated chloride conductance, EAAT5 exhibits low glutamate transport rates, but a large uncoupled chloride conductance. Mouse EAAT5 was expressed in mammalian cells und studied through whole-cell patch clamp recordings. EAAT5 exhibits uptake currents below the resolution limit. However, we observed significant anion currents in the absence as well as in the presence of glutamate, which differ in time- and voltage dependence from anion currents mediated by GLT1c. EAAT5 displays an apparent dissociation constant of 28.9 ± 7.9 mM L-glutamate (n = 7), at - 120 mV indicating an affinity for L-glutamate similar to EAAT4 (K_D = 16.6 ± 0.76 mM), a known high-affinity, low-capacity EAAT transporter. In contrast, variation of sodium concentrations at -120 mV and saturating glutamate concentrations reveal a relatively low sodium affinity of EAAT5 (K_D = 42.2 ± 9.2 mM) compared to EAAT4 (K_D = 3.9 ± 0.3 mM) and GLT1c (K_D = 26.9 ± 12.9 mM). EAATs are assembled as trimers and colocalization and functional differences of EAAT5 and GLT1c raise the question whether these transporters form hetero-oligomers with novel functions in retinal neurons. However, pull-down experiments did not reveal any evidence for direct interaction of EAAT5 and GLT1c. We conclude that in photoreceptor axon terminals high affinity low capacity and low affinity high capacity glutamate transporters co-exist without indication for functional interactions.