The Progestin Receptor Interactome in the Female Mouse Hypothalamus: Interactions with Synaptic Proteins Are Isoform Specific and Ligand Dependent.
The Progestin Receptor Interactome in the Female Mouse Hypothalamus: Interactions with Synaptic Proteins Are Isoform Specific and Ligand Dependent.
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DOI：
10.1523/eneuro.0272-17.2017
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发表时间：
2017-09
期刊：
影响因子：
3.4
通讯作者：
Tetel MJ
中科院分区：
文献类型：
--
作者：
Acharya KD;Nettles SA;Sellers KJ;Im DD;Harling M;Pattanayak C;Vardar-Ulu D;Lichti CF;Huang S;Edwards DP;Srivastava DP;Denner L;Tetel MJ
孕激素与脑内孕激素受体（PR）亚型PR- a和PR- b结合，影响发育、女性生殖、焦虑和压力。激素激活的pr与多种蛋白质结合形成功能复合物。在本研究中，利用谷胱甘肽s转移酶标记的小鼠PR- a和PR- b进行亲和下拉实验，从雌性小鼠下丘脑中分离出与PR相关的蛋白质。利用互补蛋白质组学方法、逆相蛋白阵列（RPPA）和质谱法，我们鉴定出下丘脑中与PR以配体依赖性和异构体特异性方式相互作用的蛋白，并通过Western blot进行了证实。突触蛋白，包括synapsin- 1和synapsin-II，与激动剂结合的PR亚型相互作用，表明这两种亚型在突触可塑性中起作用。为了进一步支持，synaptogyrin-III和synapsin-III分别与PR-A和PR-B相关。PR还与激酶相互作用，包括c-Src、mTOR和MAPK1，证实磷酸化是PR在大脑中快速作用的一个完整过程。PR与转录调控中的作用一致，以配体特异性和异构体依赖性的方式与转录因子和共激活因子相关。有趣的是，两种PR亚型都与能量稳态的关键调节因子fox01相关，这表明PR在能量代谢中的新作用。由于该PR相互作用组中许多已确定的蛋白是突触蛋白，因此我们验证了孕激素在突触可塑性中起作用的假设。事实上，黄体酮通过增加突触素- 1阳性突触，在大鼠初级皮层神经元培养中增强了突触密度。这种新颖的RPPA和质谱结合可以鉴定PR在突触重塑和能量稳态中的作用，并揭示了孕激素在脑功能和疾病中的独特作用。
Progestins bind to the progestin receptor (PR) isoforms, PR-A and PR-B, in brain to influence development, female reproduction, anxiety, and stress. Hormone-activated PRs associate with multiple proteins to form functional complexes. In the present study, proteins from female mouse hypothalamus that associate with PR were isolated using affinity pull-down assays with glutathione S-transferase–tagged mouse PR-A and PR-B. Using complementary proteomics approaches, reverse phase protein array (RPPA) and mass spectrometry, we identified hypothalamic proteins that interact with PR in a ligand-dependent and isoform-specific manner and were confirmed by Western blot. Synaptic proteins, including synapsin-I and synapsin-II, interacted with agonist-bound PR isoforms, suggesting that both isoforms function in synaptic plasticity. In further support, synaptogyrin-III and synapsin-III associated with PR-A and PR-B, respectively. PR also interacted with kinases, including c-Src, mTOR, and MAPK1, confirming phosphorylation as an integral process in rapid effects of PR in the brain. Consistent with a role in transcriptional regulation, PR associated with transcription factors and coactivators in a ligand-specific and isoform-dependent manner. Interestingly, both PR isoforms associated with a key regulator of energy homeostasis, FoxO1, suggesting a novel role for PR in energy metabolism. Because many identified proteins in this PR interactome are synaptic proteins, we tested the hypothesis that progestins function in synaptic plasticity. Indeed, progesterone enhanced synaptic density, by increasing synapsin-I–positive synapses, in rat primary cortical neuronal cultures. This novel combination of RPPA and mass spectrometry allowed identification of PR action in synaptic remodeling and energy homeostasis and reveals unique roles for progestins in brain function and disease.
使用支持向量机模型对孕酮受体相互作用的有效预测。
影响因子：
5.6
作者：
Liu JL;Peng Y;Fu YS
通讯作者：
Fu YS