期刊
JOURNAL OF NEUROSCIENCE
卷 24, 期 3, 页码 679-691出版社
SOC NEUROSCIENCE
DOI: 10.1523/JNEUROSCI.4985-03.2004
关键词
glutamate receptor; kainate receptor; trafficking; endoplasmic reticulum; patch clamp; immunofluorescence
资金
- NIMH NIH HHS [R03 MH065289, R03 MH65632] Funding Source: Medline
- NINDS NIH HHS [R01 NS44322, R01 NS044322] Funding Source: Medline
Intracellular trafficking of ionotropic glutamate receptors is regulated predominantly by determinants in the cytoplasmic C-terminal domain of the subunit proteins. Although AMPA receptors are found at the vast majority of excitatory synapses, synaptic kainate receptors exhibit a much more restricted distribution, suggesting that specific mechanisms exist for selective trafficking of these receptor proteins. In this report, we define a critical forward trafficking motif that is necessary for surface expression of the glutamate receptor 6 (GluR6) kainate receptor as well as chimeric proteins containing only the GluR6 C-terminal domain. The trafficking determinant was identified by tracking surface expression of green fluorescent protein-tagged GluR6 receptors with confocal immunofluorescence in COS-7 cells and cultured neurons and patch-clamp electrophysiology in human embryonic kidney 293 cells. Serial truncation and alanine site mutagenesis of the GluR6 subunit C terminus localized the critical motif to a seven amino acid stretch of predominantly basic residues. Alanine mutation of the trafficking motif reduced kainate receptor current amplitudes by >90% and resulted in retention of the mutated receptors in the endoplasmic reticulum. This forward trafficking domain is the first such identified for kainate receptors.
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