The opioid ligand binding of human μ-opioid receptor is modulated by novel splice variants of the receptor
The opioid ligand binding of human μ-opioid receptor is modulated by novel splice variants of the receptor
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DOI：
10.1016/j.bbrc.2006.03.084
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发表时间：
2006-05-19
影响因子：
3.1
通讯作者：
Loh, HH
中科院分区：
文献类型：
--
作者：
Choi, HS;Kim, CS;Loh, HH
吗啡和吗啡样药物（如海洛因）的药理作用主要通过β-阿片受体（莫尔）介导。有人提出，莫尔的功能多样性可能与莫尔基因的选择性剪接有关。尽管已经报道了许多莫尔mRNA剪接变体，但它们的生物学功能一直存在争议。在这项研究中，两个新的剪接变异体的人类莫尔基因被发现。剪接变体I和2（这里称为SV 1和SV 2）保留了内含子I的不同部分。在体外翻译的SV 1和SV 2产生的蛋白质与预测的分子量。剪接变体蛋白与野生型莫尔-1的第一个跨膜结构域相同，但在第一个胞内环结构域之后不同。RT-PCR证实hMOR的SV 1和SV 2存在于人神经母细胞瘤NMB细胞和人全脑中。在受体结合试验中，表达SV 1和SV 2的细胞不显示与[3 H]二丙诺啡的结合。莫尔的形成。SV 1和莫尔。SV 2异源二聚体通过免疫共沉淀和莫尔与剪接变体之间的生物发光共振能量转移来证明。MOR-GFP和SV-DsRed基因共转染显示，莫尔和SV蛋白共定位于细胞质膜。在表达人莫尔基因的NMB细胞中，SV 1或SV 2的转染降低了内源性莫尔的结合活性。这些数据支持SV 1和SV 2蛋白作为人β-阿片受体的可能生物调节剂的潜在作用。(c)2006年爱思唯尔公司All rights reserved.
The pharmacological actions of morphine and morphine-like drugs, such as heroin, mediate primarily through the p-opioid receptor (MOR). It has been proposed that the functional diversity of MOR may be related to alternative splicing of the MOR gene. Although a number of MOR mRNA splice variants have been reported, their biological function has been controversial. In this study, two novel splice variants of the human MOR gene were discovered. Splice variants I and 2 (here called the SV1 and SV2) retain different portions of intron I. In vitro translation of SV1 and SV2 produced proteins with the predicted molecular weights. The splice variant proteins were identical to the wild-type MOR-1 up to the first transmembrane domains, but were different after the first intracellular loop domains. SV1 and SV2 of hMOR were present in human neuroblastoma NMB cells and human whole brain confirmed by RT-PCR. In a receptor binding assay, cells expressing the SV1 and SV2 do not exhibit binding to [3 H]diprenorphine. The formations of MOR . SV1 and MOR. SV2 heterodimers were demonstrated by co-immunoprecipitation and bioluminescence resonance energy transfer between MOR and splice variants. Co-transfection of MOR-GFP and SV-DsRed gene showed that MOR and SV protein co-localized at the cytoplasmic membrane. In NMB cells expressing human MOR gene, transfection of SV1 or SV2 reduced binding activity of the endogenous MOR. These data support a potential role of SV1 and SV2 proteins as possible biological modulator of human p-opioid receptor. (c) 2006 Elsevier Inc. All rights reserved.