Integrative Proteomic and Phosphoproteomic Analyses of Granulosa Cells During Follicular Atresia in Porcine.
Integrative Proteomic and Phosphoproteomic Analyses of Granulosa Cells During Follicular Atresia in Porcine.
复制标题
猪滤泡闭锁期间颗粒细胞的综合蛋白质组学和磷酸化蛋白质组学分析
DOI：
10.3389/fcell.2020.624985
复制
发表时间：
2020
影响因子：
5.5
通讯作者：
Zeng S
中科院分区：
文献类型：
--
作者：
Yang F;Liu Q;Chen Y;Ye H;Wang H;Zeng S
卵巢卵泡闭锁是一种自然的生理过程，但其发生机制尚不完全清楚。本研究采用TMT标记、磷酸肽富集和LC-MS/MS分析方法，对猪正常卵泡（H）、轻度闭锁卵泡（SA）和闭锁卵泡（A）的颗粒细胞（GC）进行了定量蛋白质组学和磷酸蛋白质组学分析。总共对6，201种蛋白质进行了定量，并对1，760种蛋白质的4，723个磷酸化位点进行了定量。总共，在H/SA和H/A中分别鉴定了24种（11种上升，13种下降）和50种（29种上升，21种下降）具有倍数变化（FC）> 5的蛋白质。此外，还有20个（H/SA，up）和39个（H/A，up）磷酸化位点，FC > 7，可作为区分不同质量类别卵泡的潜在生物标志物。Western印迹和免疫荧光证实了蛋白质组学分析的可靠性。一些关键蛋白质（例如，MIF、β连环蛋白、整联蛋白β2）、磷酸化位点（例如，半胱天冬酶6的S76，核纤层蛋白A/C的S22和S636），途径（例如，细胞凋亡，肌动蛋白细胞骨架途径的调节），转录因子（例如，STAT 5A、FOXO 1和BCLAF 1）和激酶（例如，进一步分析了参与闭锁过程的4种蛋白（PI 3 K、CDK 5、CDK 12、AKT 3）的差异表达蛋白（DEPs）和磷酸化蛋白（DEPPs）。总而言之，本研究的蛋白质组学和磷酸蛋白质组学分析和功能研究全面分析了卵泡闭锁期间蛋白质表达和磷酸化的动态变化，并对这一过程的调控提供了一些新的解释。
Ovarian follicular atresia is a natural physiological process; however, the mechanism is not fully understood. In this study, quantitative proteomic and phosphoproteomic analyses of granulosa cells (GC) in healthy (H), slightly atretic (SA), and atretic follicles (A) of porcine were performed by TMT labeling, enrichment of phosphopeptides and LC-MS/MS analysis. In total, 6,201 proteins were quantified and 4,723 phosphorylation sites of 1,760 proteins were quantified. In total, 24 (11 up, 13 down) and 50 (29 up, 21 down) proteins with a fold change (FC) > 5 were identified in H/SA and H/A, respectively. In addition, there were 20 (H/SA, up) and 39 (H/A, up) phosphosites with an FC > 7, that could serve as potential biomarkers for distinguishing different quality categories of follicles. Western blotting and immunofluorescence confirmed the reliability of the proteomic analysis. Some key proteins (e.g., MIF, beta catenin, integrin β2), phosphosites (e.g., S76 of caspase6, S22 and S636 of lamin A/C), pathways (e.g., apoptosis, regulation of actin cytoskeleton pathway), transcription factors (e.g., STAT5A, FOXO1, and BCLAF1), and kinases (e.g., PBK, CDK5, CDK12, AKT3) involved in atresia process were revealed via further analysis of the differentially expressed proteins (DEPs) and phosphorylated proteins (DEPPs). Collectively, the proteomic and phosphoproteomic profiling and functional research in the current study comprehensively analyzed the dynamic changes in protein expression and phosphorylation during follicular atresia and provided some new explanations regarding the regulation of this process.
登录
查看更多内容
影响因子：
5.4
作者：
Freeman TC;Ivens A;Baillie JK;Beraldi D;Barnett MW;Dorward D;Downing A;Fairbairn L;Kapetanovic R;Raza S;Tomoiu A;Alberio R;Wu C;Su AI;Summers KM;Tuggle CK;Archibald AL;Hume DA
通讯作者：
Hume DA
DOI：
10.1084/jem.190.10.1375
发表时间：
1999-11-15
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Hudson JD;Shoaibi MA;Maestro R;Carnero A;Hannon GJ;Beach DH
通讯作者：
Beach DH
DOI：
10.1615/critreveukargeneexpr.v9.i3-4.130
发表时间：
1999-01-01
影响因子：
1.6
作者：
Goldberg, M;Harel, A;Gruenbaum, Y
通讯作者：
Gruenbaum, Y
影响因子：
4.4
作者：
Hatzirodos N;Irving-Rodgers HF;Hummitzsch K;Harland ML;Morris SE;Rodgers RJ
通讯作者：
Rodgers RJ
影响因子：
14.9
作者：
Hu H;Miao YR;Jia LH;Yu QY;Zhang Q;Guo AY
通讯作者：
Guo AY