Turnover analysis of N-methyl-D-aspartate receptor subunit NR1 protein in PC12 cells.
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.
Vazhappilly
Rema
R
Biotechnology Research Institute and Department of Biology, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, SAR, Hong Kong, China.
Sucher
Nikolaus J
NJ
D017398:Alternative Splicing / Q000502:physiology*; D000818:Animals; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology; D002490:Central Nervous System / Q000378:metabolism*; D003513:Cycloheximide / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology; D018698:Glutamic Acid / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000378:metabolism / Q000494:pharmacology; D007150:Immunohistochemistry; D009474:Neurons / Q000378:metabolism*; D016716:PC12 Cells; D014176:Protein Biosynthesis / Q000502:physiology; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics* / Q000378:metabolism*; D009435:Synaptic Transmission / Q000502:physiology*
C409466:NR1 NMDA receptor; D020033:Protein Isoforms; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D016194:Receptors, N-Methyl-D-Aspartate; D018698:Glutamic Acid; D003513:Cycloheximide; D006026:Glycoside Hydrolases