Protein chemical characterization and immunocytochemical localization of the NMDA receptor subunit NMDA R1.
Protein chemical characterization and immunocytochemical localization of the NMDA receptor subunit NMDA R1.
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DOI：
10.1016/s0021-9258(18)41579-7
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发表时间：
1993-10
期刊：
影响因子：
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通讯作者：
N. Brose;G. Gasic;D. Vetter;J. Sullivan;S. Heinemann
中科院分区：
文献类型：
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作者：
N. Brose;G. Gasic;D. Vetter;J. Sullivan;S. Heinemann
在大鼠中枢神经系统中，编码N-甲基-D-天冬氨酸受体亚基R1的mRNA是该谷氨酸受体亚型的克隆亚基mRNA中分布最广泛的。N-甲基-D-天冬氨酸R1 mRNA表达量非常丰富，N-甲基-D-天冬氨酸R1共表达对于所有其他克隆的N-甲基-D-天冬氨酸受体亚基的功能表达是必需的。因此，R1亚基可能是所有已知的N-甲基-D-天冬氨酸受体在大鼠脑中的一个重要组成部分。通过采用序列特异性多克隆抗体，我们表明，大鼠脑N-甲基-D-天冬氨酸R1，以及重组表达的受体蛋白，具有116 kDa的表观分子量在十二烷基硫酸钠聚丙烯酰胺凝胶电泳。受体蛋白质是高度糖基化的。它特异性地定位于中枢神经系统，并且在大脑皮层的亚细胞分级分离后与突触膜共富集。突触膜蛋白的化学交联显示N-甲基-D-天冬氨酸R1蛋白是分子量为730 kDa的受体蛋白复合物的一部分。通过使用免疫细胞化学方法，我们证明了一个广泛的，但不同的分布N-甲基-D-天冬氨酸R1在大鼠大脑的神经元，在大脑皮层的某些层，在海马和齿状回，以及在小脑突出的免疫染色。
In the rat central nervous system, the mRNA encoding the N-methyl-D-aspartate receptor subunit R1 is the most ubiquitously distributed among the cloned subunit mRNAs of this glutamate receptor subtype. The N-methyl-D-aspartate R1 mRNA is very abundantly expressed and N-methyl-D-aspartate R1 coexpression is necessary for functional expression of all other cloned N-methyl-D-aspartate receptor subunits. Therefore, the R1 subunit is likely to be an essential component of all known N-methyl-D-aspartate receptors in rat brain. By employing sequence specific polyclonal antibodies, we demonstrate that rat brain N-methyl-D-aspartate R1, as well as recombinantly expressed receptor protein, has an apparent molecular mass of 116 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis. The receptor protein is heavily glycosylated. It is specifically localized to the central nervous system, and it co-enriches with synaptic membranes upon subcellular fractionation of the cerebral cortex. Chemical cross-linking of synaptic membrane proteins shows that the N-methyl-D-aspartate R1 protein is part of a receptor protein complex with a molecular mass of 730 kDa. By using immunocytochemical methods, we demonstrate a widespread but distinct distribution of N-methyl-D-aspartate R1 in neurons of the rat brain, with prominent immunostaining in certain layers of the cerebral cortex, in the hippocampus and dentate gyrus, as well as in the cerebellum.