Cannabinoid receptor interacting protein suppresses agonist-driven CB(1) receptor internalization and regulates receptor replenishment in an agonist-biased manner.
Cannabinoid receptor interacting protein suppresses agonist-driven CB(1) receptor internalization and regulates receptor replenishment in an agonist-biased manner.
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DOI：
10.1111/jnc.13767
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发表时间：
2016-11
影响因子：
4.7
通讯作者：
Howlett AC
中科院分区：
文献类型：
--
作者：
Blume LC;Leone-Kabler S;Luessen DJ;Marrs GS;Lyons E;Bass CE;Chen R;Selley DE;Howlett AC
大麻素受体相互作用蛋白1a（CRIP 1a）是CB 1受体（CB 1 R）远端C-末端相关蛋白，其通过G蛋白调节CB 1 R信号传导，并且CB 1 R下调但不脱敏（Cell Signal.，27，716-726; Mol.药理学、87，747-765）。在这项研究中，我们确定了CRIP 1a参与CB 1 R质膜运输。为了跟踪激动剂和拮抗剂对细胞表面CB 1 R的影响，我们利用遗传上均质的克隆神经元细胞系N18 TG 2，其内源性表达CB 1 R和CRIP 1a，并表现出良好的特征性内源性大麻素信号传导系统。我们开发了稳定的CRIP 1a过表达和CRIP 1a-siRNA沉默敲低克隆，以研究CRIP 1a对CB 1 R质膜表达的基因剂量效应。结果表明，CP 55940或WIN 55212 -2（10 nM，5 min）通过发动蛋白和网格蛋白依赖性过程减少细胞表面CB 1 R，并且这通过CRIP 1a过表达而减弱。CP 55940介导的细胞表面CB 1 R损失之后，表面受体的环己酰亚胺敏感性恢复（30-120分钟），表明需要新的蛋白质合成。相反，WIN 55212 -2介导的细胞表面CB 1 R仅在CRIP 1a敲低细胞中恢复。CRIP 1a表达水平的变化不影响利莫那班（10 nM）介导的细胞表面CB 1 R的瞬时增加，这被认为是利莫那班对“非激动剂驱动”内化作用的结果。这些研究证明了CRIP 1a在激动剂驱动的CB 1 R细胞表面调节中的新作用，假定通过两种机制发生：1）减弱激动剂介导的内化，但在不存在外源激动剂的情况下不是这样，和2）从头合成的受体向细胞表面的偏向激动剂依赖性运输。
Cannabinoid receptor interacting protein 1a (CRIP1a) is a CB1 receptor (CB1R) distal C-terminus-associated protein that modulates CB1R signaling via G proteins, and CB1R down-regulation but not desensitization ( Cell Signal., 27, 716–726; Mol. Pharmacol., 87, 747–765). In this study, we determined the involvement of CRIP1a in CB1R plasma membrane trafficking. To follow the effects of agonists and antagonists on cell surface CB1Rs, we utilized the genetically homogeneous cloned neuronal cell line N18TG2, which endogenously expresses both CB1R and CRIP1a, and exhibits a well-characterized endocannabinoid signaling system. We developed stable CRIP1a-over-expressing and CRIP1a-siRNA-silenced knockdown clones to investigate gene dose effects of CRIP1a on CB1R plasma membrane expression. Results indicate that CP55940 or WIN55212-2 (10 nM, 5 min) reduced cell surface CB1R by a dynamin- and clathrin-dependent process, and this was attenuated by CRIP1a over-expression. CP55940-mediated cell surface CB1R loss was followed by a cycloheximide-sensitive recovery of surface receptors (30–120 min), suggesting the requirement for new protein synthesis. In contrast, WIN55212-2-mediated cell surface CB1Rs recovered only in CRIP1a knockdown cells. Changes in CRIP1a expression levels did not affect a transient rimonabant (10 nM)-mediated increase in cell surface CB1Rs, which is postulated to be as a result of rimonabant effects on ‘non-agonist-driven’ internalization. These studies demonstrate a novel role for CRIP1a in agonist-driven CB1R cell surface regulation postulated to occur by two mechanisms: 1) attenuating internalization that is agonist-mediated, but not that in the absence of exogenous agonists, and 2) biased agonist-dependent trafficking of de novo synthesized receptor to the cell surface.
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动力蛋白2是巨噬细胞中吞噬作用所必需的。
影响因子：
15.3
作者：
Gold, E S;Underhill, D M;Morrissette, N S;Guo, J;McNiven, M A;Aderem, A
通讯作者：
Aderem, A
影响因子：
--
作者：
Croise, Pauline;Estay-Ahumada, Catherine;Ory, Stephane
通讯作者：
Ory, Stephane
影响因子：
2.9
作者：
Keren, O;Sarne, Y
通讯作者：
Sarne, Y
通过滥交磷酸化依赖性机制调节CB1大麻素受体内在化。
影响因子：
4.7
作者：
Daigle TL;Kwok ML;Mackie K
通讯作者：
Mackie K
影响因子：
3.6
作者：
McDonald, Neil A.;Henstridge, Christopher M.;Irving, Andrew J.
通讯作者：
Irving, Andrew J.