D2 dopamine receptor expression and trafficking is regulated through direct interactions with ZIP.
D2 dopamine receptor expression and trafficking is regulated through direct interactions with ZIP.
复制标题
DOI：
10.1111/j.1471-4159.2008.05348.x
复制
发表时间：
2008-07
影响因子：
4.7
通讯作者：
Sibley DR
中科院分区：
文献类型：
--
作者：
Kim OJ;Ariano MA;Namkung Y;Marinec P;Kim E;Han J;Sibley DR
我们使用酵母双杂交系统将蛋白激酶 C-ζ 相互作用蛋白 (ZIP) 鉴定为 D2 多巴胺受体 (DAR) 的新型相互作用蛋白。通过使用 D2 DAR 的第三个细胞内环作为诱饵筛选大鼠脑 cDNA 文库，鉴定了这种相互作用。分离了编码 ZIP 的部分长度 cDNA，并表征为与 D2 DAR 的第三个细胞内环特异性相互作用，但不与其他 DAR 亚型的第三个细胞内环相互作用。通过在哺乳动物细胞中表达全长 ZIP 和 D2 DAR 蛋白并证明它们可以免疫共沉淀，获得了 ZIP-D2 DAR 相互作用的生化证实。我们进一步表明 ZIP 和 D2 DAR 可以从内源性脑组织中进行免疫共沉淀。免疫组织化学分析进一步表明，ZIP 和 D2 DAR 广泛共定位于不同大脑区域的众多神经元中。 ZIP 以三种不同长度的蛋白质亚型存在，它们源自选择性 RNA 剪接。发现所有三种亚型均与 D2 DAR 相互作用，从而将受体相互作用结构域界定在 ZIP 的 38 个残基内。从功能上讲，ZIP 的过度表达被发现会导致 D2 DAR 表达减少，同时受体对 cAMP 积累的调节也相应减少。共聚焦显微镜显示，ZIP 过度表达还会导致 D2 DAR 蛋白在溶酶体区室中的细胞内积累。这些结果表明，ZIP 可以与 D2 DAR 物理相互作用，导致细胞内向溶酶体的运输增加，随后受体表达和功能下调。
We have used the yeast two-hybrid system to identify protein kinase C-ζ interacting protein (ZIP) as a novel interacting protein for the D2 dopamine receptor (DAR). This interaction was identified by screening a rat brain cDNA library using the third intracellular loop of the D2 DAR as bait. A partial-length cDNA encoding ZIP was isolated and characterized as specifically interacting with the third intracellular loop of the D2 DAR, but not with the third intracellular loops of other DAR subtypes. Biochemical confirmation of the ZIP–D2 DAR interaction was obtained by expressing the full-length ZIP and D2 DAR proteins in mammalian cells and demonstrating that they could be co-immunoprecipitated. We further showed that ZIP and the D2 DAR could be co-immunoprecipitated from endogenous brain tissues. Immunohistochemical analyses further revealed that ZIP and the D2 DAR were extensively co-localized within numerous neurons in various brain regions. ZIP exists as three protein isoforms of varying length, which are derived from alternative RNA splicing. All three isoforms were found to interact with the D2 DAR, which allowed for the delineation of the receptor interacting domain to within 38 residues of ZIP. Functionally, over-expression of ZIP was found to result in decreased expression of the D2 DAR with a corresponding decrease in receptor modulation of cAMP accumulation. Confocal microscopy revealed that ZIP over-expression also lead to an intracellular accumulation of D2 DAR protein in lysosome compartments. These results suggest that ZIP can physically interact with the D2 DAR leading to increased intracellular trafficking to lysosomes with subsequent down-regulation of receptor expression and function.
登录
查看更多内容
影响因子：
2.8
作者：
Neve, KA;Seamans, JK;Trantham-Davidson, H
通讯作者：
Trantham-Davidson, H
DOI：
10.1016/s0169-328x(99)00093-5
发表时间：
1999-06-08
期刊：
MOLECULAR BRAIN RESEARCH
影响因子：
--
作者：
Nakaso, K;Kitayama, M;Yamada, K
通讯作者：
Yamada, K
影响因子：
64.8
作者：
MONSMA, FJ;MCVITTIE, LD;SIBLEY, DR
通讯作者：
SIBLEY, DR
影响因子：
4.8
作者：
Croci, C;Brandstätter, JH;Enz, R
通讯作者：
Enz, R
影响因子：
2.5
作者：
McNaught, KSP;Jenner, P
通讯作者：
Jenner, P