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KMID : 0578319950050050436
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Molecules and Cells
1995 Volume.5 No. 5 p.436 ~ p.441
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Immunological Evidence for the enrichment of N-methyl-D-aspartase Recpetor Subunit 2A in the Rat Postsynaptic Density
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Moon, Il Soo
Jin, Deuk Hee/Ko, Bok Hyun
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Abstract
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The synapse is a special structure where communication between neurons takes place. One of the postsynaptic structures that is thought to have a regulatory role in synaptic transmission is the postsynaptic density (PSD). As an effort to understand the subcellular localization of N-methyl-D-aspartate (NMDA) receptors, which play an important role in synaptic plasticity, we carried out immunological experiments to study whether NMDA receptor subunit 2A (NR2A) is present in the PSD and enrichd there. In oder to generate NR2A-specific antibody, we amplified a segment of NR2A gene by polymerase chain reaction and subcloned the fragment into a bacterial expression vector pGEX2T. We expressed the NR2A segment as a glutathione-S-transferase fusion protein in Escherichia coli, and made anti-NR2A antibodies using the fusion protein. Immunoblot analyses revealed that the molecular size of NR2A is 180 kDa, that NR2A contains a similar amount of N-linked oligosaccharide, and that NR2A is highly enriched in the forebrain PSD fraction. Our data indicate that NR2A is specifically associated with PSDs reinforcing the idea that the PSD is a key synaptic regulator.
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