Abstract
Publisher Summary This chapter describes a biochemical procedure to obtain a highly purified glycine transport activity with GLYT2-like properties from pig brain stem. The main advantage of this procedure is to provide the transporter in its native state, what readily permits the study of its biochemical, structural, and functional properties. Glycine acts as an inhibitory neurotransmitter in the central nervous system (CNS) of vertebrates, mainly in the spinal cord and the brain stem. An additional role of glycine is the potentiation of glutamate excitatory action on postsynaptic N-methyl-D-aspartate (NMDA) receptors. The reuptake of glycine into presynaptic nerve terminals or neighboring glial cells provides one way of clearing the neurotransmitter from the extracellular space and constitutes an efficient mechanism by which its postsynaptic action can be terminated. This process is carried out by a transport system that actively accumulates glycine and that is energized by the electrochemical gradient of sodium. The purification of glycine transporter involves the solubilization of the transporter protein from plasma membrane vesicles followed by three chromatographic steps on phenyl-Sepharose, wheat germ agglutinin-Sepharose and hydroxylapatite columns, and a final sucrose density gradient fractionation.
Published Version
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