Zhou Yuxin, Qiu Liyan, Sterpka Ashley, Wang Haiying, Chu Feixia, Chen Xuanmao
Department of Molecular, Cellular and Biomedical Sciences, University of New Hampshire, Durham, NH, United States.
Department of Statistics, University of Connecticut, Storrs, CT, United States.
Front Cell Neurosci. 2019 Feb 13;13:34. doi: 10.3389/fncel.2019.00034. eCollection 2019.
Type III adenylyl cyclase (AC3, ) is predominantly enriched in neuronal primary cilia throughout the central nervous system (CNS). Genome-wide association studies in humans have associated with major depressive disorder and autistic spectrum disorder, both of which exhibit sexual dimorphism. To date, it is unclear how AC3 affects protein phosphorylation and signal networks in central neurons, and what causes the sexual dimorphism of autism. We employed a mass spectrometry (MS)-based phosphoproteomic approach to quantitatively profile differences in phosphorylation between inducible AC3 knockout (KO) and wild type (WT), male and female mice. In total, we identified 4,655 phosphopeptides from 1,756 proteins, among which 565 phosphopeptides from 322 proteins were repetitively detected in all samples. Over 46% phosphopeptides were identified in at least three out of eight biological replicas. Comparison of AC3 KO and WT datasets revealed that phosphopeptides with motifs matching proline-directed kinases' recognition sites had a lower abundance in the KO dataset than in WTs. We detected 14 phosphopeptides restricted to WT dataset (i.e., ) and 35 exclusively in KOs (i.e., ). Moreover, 95 phosphopeptides (out of 90 proteins) were identified only in female dataset and 26 only in males. Label-free MS spectrum quantification using Skyline further identified phosphopeptides that had higher abundance in each sample group. In total, 204 proteins had sex-biased phosphorylation and 167 of them had increased expression in females relative to males. Interestingly, among the 204 gender-biased phosphoproteins, 31% were found to be associated with autism, including . Therefore, this study also provides the first phosphoproteomics evidence suggesting that gender-biased post-translational phosphorylation may be implicated in the sexual dimorphism of autism.
III型腺苷酸环化酶（AC3）在整个中枢神经系统（CNS）的神经元初级纤毛中高度富集。全基因组关联研究表明，AC3与重度抑郁症和自闭症谱系障碍有关，这两种疾病都表现出性别差异。迄今为止，尚不清楚AC3如何影响中枢神经元中的蛋白质磷酸化和信号网络，以及自闭症的性别差异是由什么引起的。我们采用基于质谱（MS）的磷酸化蛋白质组学方法，定量分析了可诱导的AC3基因敲除（KO）和野生型（WT）雄性和雌性小鼠之间磷酸化的差异。我们总共从1756种蛋白质中鉴定出4655个磷酸肽段，其中来自322种蛋白质的565个磷酸肽段在所有样本中均被重复检测到。超过46%的磷酸肽段在八个生物学重复中的至少三个中被鉴定出来。AC3基因敲除组和野生型组数据集的比较表明，与脯氨酸定向激酶识别位点匹配基序的磷酸肽段在基因敲除组数据集中的丰度低于野生型组。我们检测到14个仅在野生型组数据集中存在的磷酸肽段（即）和35个仅在基因敲除组中存在的磷酸肽段（即）。此外，在雌性数据集中仅鉴定出95个磷酸肽段（来自90种蛋白质），在雄性数据集中仅鉴定出26个。使用Skyline进行的无标记MS谱定量进一步鉴定了每个样本组中丰度较高的磷酸肽段。总共有204种蛋白质存在性别偏向性磷酸化，其中167种在雌性中的表达相对于雄性有所增加。有趣的是，在这204种性别偏向性磷酸化蛋白质中，发现31%与自闭症有关，包括。因此，本研究还提供了首个磷酸化蛋白质组学证据，表明性别偏向性翻译后磷酸化可能与自闭症的性别差异有关。
