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VGLUT1 is expressed in a subset of glutamate neurons and transports glutamate into native synaptic vesicles from the brain, exhibiting a conductance for chloride that is blocked by glutamate. Vesicular glutamate transport has a substantially lower apparent affinity than the plasma membrane excitatory amino acid transporters. Glutamate transport by VGLUT1 is saturated with a K(m) of approximately 2 mM, in the same range as transport by synaptic vesicles. Finally, plasma membrane glutamate transporters recognize both aspartate and glutamate as substrates, whereas VGLUT1 does not recognize aspartate.