Abstract
The post-translational fate of N-methyl-D-aspartate receptor (NMDAR) subunit NR1 was characterized in PC12 cells using pulse-chase labeling, block of protein synthesis by cyclohexamide and deglycosylation by endoglycosidase H. Metabolic labeling of NR1 protein indicated a biphasic degradation of NR1 protein with half-lives of 1.6 and 16.1 h for a rapidly (78%) and a slowly (22%) degrading population. Immunoprecipitation of NR1 following the block of protein synthesis by cyclohexamide revealed that the rapidly and slowly degrading pools mainly consisted of the NR1 splice variants NR1-4a and NR1-2a. Sensitivity of NR1 protein to deglycosylation by endoglycosidase H indicated the presence of an immature form of NR1 that was retained in the endoplasmic reticulum. PC12 cells serve as a useful model for the elucidation of translational and post-translational mechanisms of NMDAR expression.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alternative Splicing / physiology*
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Animals
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Calcium Signaling / drug effects
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Calcium Signaling / physiology
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Central Nervous System / metabolism*
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Cycloheximide / pharmacology
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Endoplasmic Reticulum / metabolism
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Gene Expression Regulation / physiology
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Glutamic Acid / metabolism*
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Glycoside Hydrolases / metabolism
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Glycoside Hydrolases / pharmacology
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Immunohistochemistry
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Neurons / metabolism*
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PC12 Cells
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Protein Biosynthesis / physiology
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Protein Isoforms / genetics
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Protein Isoforms / metabolism
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Protein Synthesis Inhibitors / pharmacology
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RNA, Messenger / metabolism
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Rats
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Receptors, N-Methyl-D-Aspartate / genetics*
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Receptors, N-Methyl-D-Aspartate / metabolism*
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Synaptic Transmission / physiology*
Substances
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NR1 NMDA receptor
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Protein Isoforms
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Protein Synthesis Inhibitors
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RNA, Messenger
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Receptors, N-Methyl-D-Aspartate
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Glutamic Acid
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Cycloheximide
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Glycoside Hydrolases