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Differential surface expression and phosphorylation of the N-methyl-D-aspartate receptor subunits NR1 and NR2 in cultured hippocampal neurons.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Hall, Randy A. Soderling, Thomas R. |
| Copyright Year | 1997 |
| Abstract | The trafficking and phosphorylation of the NR1 and NR2 subunits of the N-methyl-D-aspartate-type glutamate receptor complex were studied in cultured rat hippocampal neurons. Surface expression was examined by modifying surface receptors via treatment of intact neurons with either the protease chymotrypsin or the cross-linking reagent bis(sulfosuccinimidyl)suberate, followed by quantification of anti-NR1 and anti-NR2B Western blot immunostaining. These studies revealed that only 40-50% of total NR1 immunoreactivity is found at the cell surface, as compared to more than 90% of total NR2B immunoreactivity. Metabolic labeling of the cultures with 32P revealed that NR2 subunits are highly phosphorylated under basal conditions, whereas basal phosphorylation of NR1 subunits is barely detectable. Following stimulation of the cultures with glutamate/glycine or phorbol esters, NR1 phosphorylation was found to be enhanced by 3-5-fold, whereas phosphorylation of NR2 subunits was enhanced by less than 2-fold. To determine whether the difference in the basal NR1 versus NR2 phosphorylation could be due to tyrosine phosphorylation of NR2, phosphoamino acid analyses of NR2 were performed. These analyses revealed phosphorylation on serine but not on threonine or tyrosine; immunoprecipitation and deglycosylation experiments using anti-phosphotyrosine antibodies confirmed that NR2 subunits in the primary hippocampal cultures are not detectably phosphorylated on tyrosine residues. These results demonstrate that the NR1 and NR2 subunits, which assemble into heteromeric complexes to form functional N-methyl-D-aspartate receptors, are trafficked in neurons with differential efficiency to the plasma membrane and exhibit different levels of basal versus stimulated serine phosphorylation. |
| Starting Page | 7986 |
| Ending Page | 7990 |
| Page Count | 5 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | http://www.pharm.emory.edu/rhall/rr1.pdf |
| Alternate Webpage(s) | http://pharmacology.emory.edu/hall/publications/rr1.pdf |
| Alternate Webpage(s) | http://www.pharm.emory.edu/hall/publications/rr1.pdf |
| PubMed reference number | 9020125v1 |
| Volume Number | 272 |
| Issue Number | 7 |
| Journal | The Journal of biological chemistry |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Aspartic Acid CHYMOTRYPSIN Endopeptidases Glutamate Receptor Glutamic Acid Glycine Greater Than Hippocampus (Brain) Immunoprecipitation N-Methyl-D-Aspartate Receptors N-Methylaspartate NDOR1 gene NMDA receptor 2B NR1 protein, human Phorbol Esters Phorbols Phosphotyrosine Plasma membrane Quantitation Reagents Thioctic Acid Threonine Tyrosine Phosphorylation Western Blot bis(sulfosuccinimidyl)suberate |
| Content Type | Text |
| Resource Type | Article |
