Ligand binding properties of muscarinic acetylcholine receptor subtypes (m1-m5) expressed in baculovirus-infected insect cells.
Ligand binding properties of muscarinic acetylcholine receptor subtypes (m1-m5) expressed in baculovirus-infected insect cells.
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在杆状病毒感染的昆虫细胞中表达的毒蕈碱乙酰胆碱受体亚型（m1-m5）的配体结合特性。
DOI：
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发表时间：
1995
影响因子：
3.5
通讯作者：
Tatsuya Haga
中科院分区：
文献类型：
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作者：
Guo Zheng Dong;Kimihiko Kameyama;A. Rinken;Tatsuya Haga
利用杆状病毒系统在昆虫细胞（Spodoptera frugiperda，Sf 9）中表达了5种毒蕈碱型乙酰胆碱受体亚型（m1-m5）。1升培养物可产生高达6 nmol的毒蕈碱乙酰胆碱受体; 0.3至0.6（人m1）、3至6（人m2）、2至4（大鼠m3）、1至2（大鼠m4）和0.5至1（人m5）nmol。哌仑西平、AF-DX116和hexahidrosiladifenidol分别对m1、m2和m3亚型表现出最高的亲和力，表明这些在Sf 9细胞中表达的受体保留了与哺乳动物组织或培养细胞中相同的底物特异性。在本研究中检测的32种毒蕈碱配体中，发现prifinium对m4亚型具有最高的亲和力，毛果芸香碱、氧化震颤素、McN-A343和异丙嗪对m5亚型具有最高的亲和力，尽管5种亚型之间的亲和力差异小于10倍。阿库溴铵增加了[3 H] N-甲基东莨菪碱与m2亚型的结合，但不增加m1、m4和m5亚型的结合，仅轻微增加了m3亚型的结合。类似的，但更小的影响fangchinoline和粉防己碱被发现为[3 H] N-甲基东莨菪碱结合只有m3亚型。这些影响也可能是有用的个别亚型的歧视。
Five subtypes of muscarinic acetylcholine receptors (m1-m5) have been expressed in insect cells (Spodoptera frugiperda, Sf9) using the baculovirus system. Up to 6 nmol of muscarinic acetylcholine receptors were produced by 1 liter culture; 0.3 to 0.6 (human m1), 3 to 6 (human m2), 2 to 4 (rat m3), 1 to 2 (rat m4) and 0.5 to 1 (human m5) nmol. Pirenzepine, AF-DX116 and hexahidrosiladifenidol showed the highest affinity for the m1, m2 and m3 subtype, respectively, indicating that these receptors expressed in Sf9 cells retain the same substrate specificity as those in mammalian tissues or cultured cells. Among 32 kinds of muscarinic ligands examined in the present studies, prifinium was found to have the highest affinity for the m4 subtype, and pilocarpine, oxotremorine, McN-A343 and promethazine the highest affinity for the m5 subtype, although the differences in the affinities among the five subtypes were less than 10-fold. Alcuronium increased the binding of [3H]N-methylscopalamine to the m2 subtype, but not the m1, m4 and m5 subtypes and only slightly to the m3 subtype. Similar but smaller effects of fangchinoline and tetrandrine were found for [3H]N-methylscopalamine binding to only the m3 subtype. These effects may also be useful for the discrimination of individual subtypes.