摘要
为探讨κ-受体(κ-opioid receptor,KOR)和阿片受体样受体(opioid receptor like-1 receptor,ORL1 receptor)是否能够形成异源性受体二聚体,在原代培养的大鼠神经元细胞和用带有HA(hemagglutinin,血细胞凝集素)、Myc或Flag标签的KOR和ORL1质粒共同转染的中国仓鼠卵巢(CHO)细胞、人胚肾上皮(HEK293)细胞上,采用免疫荧光和免疫共沉淀的方法,研究KOR和ORL1之间的共定位以及是否存在相互作用。结果表明:在原代培养的海马和皮质神经元上,KOR和ORL1的免疫荧光在细胞膜上有重叠。同样,在HA-KOR和Myc-ORL1共同瞬时转染的CHO和HEK293细胞上也有类似的发现。另外,在共同表达Flag-KOR和Myc-ORL1的CHO细胞裂解液中,KOR与ORL1的受体蛋白能够被相互免疫共沉淀。这些研究结果提示,作为阿片受体不同亚型的KOR和ORL1受体之间有可能存在着异源二聚体,这也为进一步解释阿片受体结构的多样性和功能的复杂性提供了新的实验依据。
This study investigates whether κ-opioid receptor and ORL1 receptor may interact to form a heterodimer.In immunofluorescence and co-immunoprecipitation experiments,differentially epitope-tagged receptors,colocalization and heterodimerization of κ-opioid receptor and ORL1 receptor were used and examined in primary culturing rat neurons,Chinese hamster ovary（CHO） or human embryonic kidney 293（HEK293） cells.The results show that fluorescence of both κ-opioid receptor and ORL1 receptor were overlapping in primary culturing hippocampal and cortical neurons.Similarly in co-expressing CHO or HEK293 cells,HA-KOR and Myc-ORL1 were almost exclusively confined to the membranes,revealing extensive colocalization.When Flag-KOR and Myc-ORL1 were co-expressing in CHO cells,heterodimerization was identified to have the ability to co-immunoprecipitate ORL1-receptors with κ-opioid receptor and vice versa.In the current study,further evidence was provided for the direct interaction of two subtypes of opioid receptors,κ-opioid receptor and ORL1-receptor,to form the heterodimerization.The finding represents the novel pharmacological mechanism for modulation of opioid receptor function as well as diversity of G protein-coupled receptors.
出处
《药学学报》
CAS
CSCD
北大核心
2011年第9期1078-1083,共6页
Acta Pharmaceutica Sinica
基金
国家高技术研究发展计划(863计划)资助项目(2005AA233040)
国家科技部"重大新药创制"科技重大专项(2009ZXJ09004-079)
