Nicotinic acetylcholine receptors of Drosophila:: three subunits encoded by genomically linked genes can co-assemble into the same receptor complex
Nicotinic acetylcholine receptors of Drosophila:: three subunits encoded by genomically linked genes can co-assemble into the same receptor complex
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DOI：
10.1046/j.0022-3042.2001.00685.x
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发表时间：
2002-01-01
影响因子：
4.7
通讯作者：
Gundelfinger, ED
中科院分区：
文献类型：
--
作者：
Chamaon, K;Smalla, KH;Gundelfinger, ED
第二个 β 样亚基 (SBD) 是果蝇烟碱乙酰胆碱受体 (nAChR) 的假定结构亚基。在这里，我们生产了针对 SBD 的特异性抗体来研究，其他 nAChR 亚基可以与果蝇神经系统受体复合物中的 SBD 共同组装。成人视叶的免疫组织化学研究表明，SBD 的分布与突触神经毡中的 α 亚基 ALS 相似。亚基 ALS、Dalpha2 和 SBD 可以通过 α-银环蛇毒素亲和层析进行共纯化。此外，抗SBD抗体共沉淀ALS和Dalpha2，反之亦然，ALS和Dalpha2抗体共免疫沉淀SBD蛋白。使用针对 ALS 和 Dalpha2 的固定化抗体进行的两步免疫亲和层析揭示了 nAChR 复合物的存在，其中包括 ALS、Dalpha2 和 SBD 作为组成部分。有趣的是，编码这三个亚基的基因似乎在果蝇基因组中第三染色体的 96 A 区域直接连锁。此外，SBD 似乎是不同受体复合物的组成部分，该复合物包括 ARID 蛋白作为额外的 β 亚基，但既不是 ALS，也不是 Dalpha2，也不是第三个 α 亚基 Dalpha3。这些发现表明果蝇烟碱受体系统相当复杂。
The second beta-like subunit (SBD) is a putative structural subunit of Drosophila melanogaster nicotinic acetylcholine receptors (nAChRs). Here we have produced specific antibodies against SBD to study, which other nAChR subunits can co-assemble with SBD in receptor complexes of the Drosophila nervous system. Immunohistochemical studies in the adult optic lobe revealed that SBD has a distribution similar to that of the alpha-subunit ALS in the synaptic neuropil. The subunits ALS, Dalpha2 and SBD can be co-purified by alpha-bungarotoxin affinity chromatography. Moreover, anti-SBD antibodies co-precipitate ALS and Dalpha2 and, vice versa, ALS and Dalpha2 antibodies co-immunoprecipitate SBD protein. Two-step immunoaffinity chromatography with immobilized antibodies against ALS and Dalpha2 revealed the existence of nAChR complexes that include ALS, Dalpha2 and SBD as integral components. Interestingly, the genes encoding these three subunits appear to be directly linked in the Drosophila genome at region 96 A of the third chromosome. In addition, SBD appears to be a component of a different receptor complex, which includes the ARID protein as an additional beta-subunit, but neither ALS nor Dalpha2 nor the third alpha-subunit Dalpha3. These findings suggest a considerable complexity of the Drosophila nicotinic receptor system.