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Proteomic Analysis of Nuclear Phosphorylated Proteins in Dairy Cow Mammary Epithelial Cells Treated with Prolactin 认领 引用
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作者 Huang Jian-guo Lu Li-min +5 位作者 Qiao Bin Liu Rong Wang Jia-li Pan Hong-bao Li Qing-zhang Gao Xue-jun 《Journal of Northeast Agricultural University(English Edition)》 CAS 2013年第3期31-39,共9页
Prolactin （PRL） is a versatile signaling molecule and regulates a variety of physiological processes, including mammary gland growth and differentiation and the synthesis of milk proteins. While PRL is known to be n... Prolactin （PRL） is a versatile signaling molecule and regulates a variety of physiological processes, including mammary gland growth and differentiation and the synthesis of milk proteins. While PRL is known to be necessary for high levels of milk protein expression, the mechanism by which the synthesis of milk proteins is stimulated at the transcript level is less known. A major modification in the transcript level is protein phosphorylation. To gain additional insights into the molecular mechanisms at the transcript level underlying PRL action on the dairy cow mammary epithelial cells （DCMECs）, nuclear phosphoproteins whose expression distinguishes proliferating regulated by PRL in DCMECs were identified. A phosphoprotein-enriched fraction from nuclear proteins was obtained by affinity chromatography, and a two-dimensional gel electrophoresis （2-DE） and matrix assisted laser desorption/ionization time of matrix-assisted laser desorption/ionizationime of flight mass spectrometry （MALDI-TOF MS） were used to identify the changes of nuclear phosphoproteins in DCMECs treated with prolactin. Seven proteins displaying~〉2-fold difference in abundance upon PRL treatment in DCMECs were identified by MALDI-TOF MS. The protein-GARS （GlyRS）, which belonged to the class-II aminoacyl-tRNA synthetase family, played a global role in the milk protein synthesis. SERPINH1 （Heat shock protein 47）, which was the first heat shock protein found to be a member of the serpin superfamily, regulated physiologic functions, such as complement activation, programmed cell death, and inflammatory processes. PRDX3, which belonged to a family of antioxidant enzymes, played an important role in scavenging intracellular reactive oxygen species （ROS）. ACTR1A, belonged to the actin family, which was associated with transport of p53 to the nucleus. Annexin A2, a Ca2＋-dependent phospholipid-binding protein, maintained the viability and cell cycle regulation of DCMECs. PSMB2 and PSMD10, which belonged to ubiquitin-proteasome system, were involved in several cellular processes, including cell cycle control, cellular stress response, intracellular signaling. This screening revealed that prolactin influenced the level of nuclear phosphoproteins in DCMECs. This result opens new avenues for the study of the molecular mechanism linked to the synthesis of milk proteins. 展开更多
关键词 nuclear phosphorylated protein prolactin 2-DE
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探讨电针对颅脑损伤大鼠认知功能及脑组织相关蛋白p-TSC2、p-p70S6K表达影响 认领 引用
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作者 王若璇 吴涛 +1 位作者 夏曾彩 黄威 《辽宁中医药大学学报》 CAS 2026年第4期35-42,共8页
目的该研究旨在探究电针对创伤性颅脑损伤(TBI)大鼠认知功能、脑组织形态结构和脑组织中磷酸化结节性硬化复合物2(p-TSC2)、磷酸化p70核糖体蛋白S6激酶(p-p70S6K)蛋白的影响。方法随机将SD雄性大鼠分为空白组、假手术组、模型组和电针... 目的该研究旨在探究电针对创伤性颅脑损伤(TBI)大鼠认知功能、脑组织形态结构和脑组织中磷酸化结节性硬化复合物2(p-TSC2)、磷酸化p70核糖体蛋白S6激酶(p-p70S6K)蛋白的影响。方法随机将SD雄性大鼠分为空白组、假手术组、模型组和电针组。使用Fenney打击法进行模型制备。电针组选取“百会、关元、曲池、合谷、足三里、涌泉”,空白组和假手术组不做其他处理。在造模后24 h进行电针干预,选用疏密波,频率1 Hz,强度1 mA,1次/d,15 min/次,连续治疗14 d。采用Morris水迷宫检测电针干预对TBI大鼠认知功能的影响;采用HE染色法观察在电针干预后的3个时间点(第3天、第7天、第14天)后,大鼠脑组织形态结构变化;采用免疫荧光染色、Western Blot实验检测在电针干预后TBI模型大鼠创伤脑组织中p-TSC2、p-p70S6K蛋白表达情况。结果在造模前、电针干预后第3天、第7天和第14天,对4组SD大鼠分别进行逃避潜伏期测试。相对于模型组,电针组逃避潜伏期在电针干预后的三个时间点呈逐渐降低的趋势(P<0.05)。电针干预第3天,模型组炎性细胞浸润,可见大量脑组织渗出液,组织空泡样变化,核固缩和核裂解明显,电针组优于模型组;电针干预第7天、第14天,与模型组相比,电针组炎症细胞减少、正常神经元增多。在电针干预后的3个时间点,电针组p-p70S6K蛋白的相对表达量相较于模型组上调(P<0.05),而p-TSC2蛋白的相对表达量下调(P<0.05)。结论电针可能通过调节自噬相关蛋白p-TSC2、p-p70S6K的表达,进一步介导细胞自噬,从而促进认知功能的恢复和对脑组织的保护作用。 展开更多
关键词 TBI 电针 AMPK/mTOR通路 磷酸化结节性硬化复合物2 磷酸化p70核糖体蛋白S6激酶 自噬
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REGULATED PHOSPHORYLATION OF THE GATA-2 DNA BINDINGPROTEIN IN ENDOTHELIAL CELLS 认领 引用
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作者 卜昕 《Journal of Pharmaceutical Analysis》 CAS 1995年第2期174-175,共2页
Endothelin-1 and a number of other genes expressd primarily in endothelial cells(EC)require a functional GATA element in their promoter region.The widely expressed zinc finger DNA binding protein GATA-2 has been chara... Endothelin-1 and a number of other genes expressd primarily in endothelial cells(EC)require a functional GATA element in their promoter region.The widely expressed zinc finger DNA binding protein GATA-2 has been characterized as the likely GATA factor which binds these GATA elements.To understand the specificity of this interaction,and to investigate the potential for regulation of GATA-2 activity,we have studied translation and post-translational modification of the GATA-2 protein. A specific antiserum immunoprecipitated a 52kDa GATA-2 protein from [35-S] methionine-labeled EC,as well as a wide variety of cultured human cell lines which express GATA-2 mRNA. Immunoprecipitation experiments with [32-P]-orthophosphate labeled cells indicated that GATA-2 is similarly phosphorylated in EC and non-EC lines. Thus the apparent cell-specific activity of this transcription factor is not regulated by translation or phosphorylation, and must derive from the interaction of GATA-2 with other nuclear proteins in the EC.Further studies investigated the potential regulation of GATA-2 phosphorylation in EC. Phosphoamino acid analysis indicated that GATA-2 is phosphorylated on serine and threonine residues in EC.The hasal phosphorylation of GATA-2 was rapidly and markedly increased when EC were treated with calcium ionophore A23187, while phorbol ester and forskolin had no effect.Phosphopeptide map analysis showed that A23187 induced phosphorylation of at least two additional sites in GATA-2.Gel shift assays employing nuclear extracts isolated from EC that had been treated with A23187 had a different DNA binding pattern when compared to control.This regulated phosphorylation of GATA-2 may provide a signaling pathway for hormonal regulation of endothelial cell genes such as endothelin-1 which alter their rate of transcription in response to increased intracellular calcium. 展开更多
关键词 GATA-2 endothelial cell endothelin-1 phosphorylation DNA binding protein
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Pathogen-induced H2O2triggers phosphorylation of the RNA-binding protein RRM2R to suppress OsMAPKKK18 intron retention for orchestrating rice immunity and growth 认领 引用 被引量：2
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作者 Xiang Lu Yibang An +26 位作者 Wenxiu Lu Jing Li Qing Xiong Miaomiao Zhu Yao He Jia Yang Yue Wang Zeming Zhang Xiaorong Gong Yulong Qian Wenyangyang Yu Yuchun Shao Huabin Zhou Xiaolin Chen Haijuan Tang Qingqing Hou Hui Shi Junjie Yin Yongyan Tang Xiaobo Zhu Yang Zhou Li Song Long Wang Min He ∙Weitao Li Jing Wang Xuewei Chen 《Molecular Plant》 SCIE CAS CSCD 2026年第1期116-133,共18页
Alternative splicing plays a vital role in plant immunity,but how plants stringently and selectively regulate alternative splicing to coordinate immunity and growth is poorly understood.Here,we report that H2O2-... Alternative splicing plays a vital role in plant immunity,but how plants stringently and selectively regulate alternative splicing to coordinate immunity and growth is poorly understood.Here,we report that H2O2-triggered phosphorylation of the RNA-binding protein RRM2R releases a retained-intron brake in OsMAPKKK18 encoding a mitogen-activated protein kinase kinase kinase to orchestrate plant immunity and growth in rice.We found that,without pathogen infection,OsMAPKKK18β transcripts containing a retained intron are subjected to nonsense-mediated mRNA decay to promote plant growth.Upon Magnaporthe oryzae pathogen invasion,H2O2activates the protein kinase OsCLK1 to phosphorylate and stabilize RRM2R,which interacts with OsRNPS1A/B to recruit spliceosome machinery and promote the production of OsMAPKKK18α transcripts without the retained intron to promote rice immunity.Furthermore,we revealed that after the endogenous H2O2is consumed or degraded by peroxidases or catalases,decreased abundance of RRM2R reduces the accumulation of OsMAPKKK18α transcripts to avoid autoimmunity in rice.Taken together,the results of our study uncover H2O2-controlled OsMAPKKK18 intron retention as a novel molecular switch for orchestrating immunity and growth in rice. 展开更多
关键词 H2O2 alternative splicing intron retention RNA-binding protein phosphorylation PTI ethylene immunity and growth
Protein tyrosine phosphatase non-receptor type 2 andinflammatory bowel disease 认领 引用 被引量：6
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作者 Marianne R Spalinger Declan F McCole +1 位作者 Gerhard Rogler Michael Scharl 《World Journal of Gastroenterology》 SCIE CAS 2016年第3期1034-1044,共11页
Genome wide association studies have associated single nucleotide polymorphisms within the gene locus encoding protein tyrosine phosphatase non-receptor type 2(PTPN2) with the onset of inflammatory bowel disease(IBD) ... Genome wide association studies have associated single nucleotide polymorphisms within the gene locus encoding protein tyrosine phosphatase non-receptor type 2(PTPN2) with the onset of inflammatory bowel disease(IBD) and other inflammatory disorders. Expression of PTPN2 is enhanced in actively inflamed intestinal tissue featuring a marked up-regulation in intestinal epithelial cells. PTPN2 deficient mice suffer from severe intestinal and systemic inflammation and display aberrant innate and adaptive immune responses. In particular, PTPN2 is involved in the regulation of inflammatory signalling cascades, and critical for protecting intestinal epithelial barrier function, regulating innate and adaptive immune responses, and finally for maintaining intestinal homeostasis. On one hand, dysfunction of PTPN2 has drastic effects on innate host defence mechanisms, including increased secretion of pro-inflammatory cytokines, limited autophagosome formation in response to invading pathogens, and disruption of the intestinal epithelial barrier. On the other hand, PTPN2 function is crucial for controlling adaptive immune functions, by regulating T cell proliferation and differentiation as well as maintaining T cell tolerance. In this way, dysfunction of PTPN2 contributes to the manifestation of IBD. The aim of this review is to present an overview of recent findings on the role of PTPN2 in intestinal homeostasis and the impact of dysfunctional PTPN2 on intestinal inflammation. 展开更多
关键词 Protein tyrosine phosphatase non-receptortype 2 Inflammatory bowel disease Chronic intestinalinflammation Barrier function Phosphorylation
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青蒿琥酯对原代慢性粒细胞白血病细胞生长抑制、凋亡诱导及bcr/abl融合基因、P210蛋白、磷酸化SHP-2蛋白表达的影响 认领 引用 被引量：4
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作者 罗曼 黄晓华 +1 位作者 古学奎 杨洪涌 《中药新药与临床药理》 CAS CSCD 北大核心 2020年第11期1289-1295,共7页
目的探讨青蒿琥酯对原代慢性粒细胞白血病细胞生长抑制、凋亡诱导及bcr/abl融合基因、P210蛋白、Src同源区2含域磷酸酶-2(SHP-2)蛋白磷酸化的影响。方法分离培养原代慢性粒细胞白血病细胞,采用不同浓度青蒿琥酯(0、5、10、20μg·mL... 目的探讨青蒿琥酯对原代慢性粒细胞白血病细胞生长抑制、凋亡诱导及bcr/abl融合基因、P210蛋白、Src同源区2含域磷酸酶-2(SHP-2)蛋白磷酸化的影响。方法分离培养原代慢性粒细胞白血病细胞,采用不同浓度青蒿琥酯(0、5、10、20μg·mL-1)进行干预,四甲基偶氮唑盐比色法(MTT)检测细胞生长抑制情况;细胞计数法检测细胞生存情况;罗丹明123染色法检测细胞线粒体膜电位的变化;电镜观察细胞超微结构的变化;流式细胞术(FCM)检测青蒿琥酯对细胞凋亡及胀亡的影响;荧光定量聚合酶链式反应(RT-PCR)检测bcr/abl融合基因mRNA表达;Western Blot检测P210、SHP-2及磷酸化SHP-2蛋白表达。结果青蒿琥酯可抑制原代慢性粒细胞白血病细胞的生长,降低细胞存活率和线粒体膜电位水平(P<0.05),且与药物浓度和时间呈现一定的依赖性;不同浓度药物和不同作用时间可导致细胞凋亡率及胀亡率明显升高(P<0.01),同时可降低bcr/abl融合基因mRNA、P210、SHP-2和磷酸化SHP-2蛋白的表达水平(P<0.05,P<0.01)。结论青蒿琥酯可通过抑制原代慢性粒细胞白血病细胞的生长,促进其凋亡和胀亡,以及降低bcr/abl融合基因mRNA、P210、SHP-2、磷酸化SHP-2蛋白在原代慢性粒细胞白血病细胞中的表达水平,达到治疗慢性粒细胞白血病(CML)的目的。 展开更多
关键词 青蒿琥酯 慢性粒细胞白血病 细胞生长 细胞凋亡 bcr/abl融合基因 P210蛋白 磷酸化SHP-2蛋白
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2型糖尿病及其并发症的翻译后修饰组学研究进展 认领 引用 被引量：2
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作者 陈秀兰 杨福全 《生物工程学报》 EI CAS CSCD 北大核心 2025年第11期4232-4249,共18页
2型糖尿病发病率和发病人数的持续攀升给社会带来了很大的经济负担。2型糖尿病的发病机制主要是胰岛β细胞的胰岛素分泌功能受损和骨骼肌、肝脏和脂肪组织的外周胰岛素抵抗。尽管已经有很多研究探索了2型糖尿病的病理机制,但是其发生和... 2型糖尿病发病率和发病人数的持续攀升给社会带来了很大的经济负担。2型糖尿病的发病机制主要是胰岛β细胞的胰岛素分泌功能受损和骨骼肌、肝脏和脂肪组织的外周胰岛素抵抗。尽管已经有很多研究探索了2型糖尿病的病理机制,但是其发生和发展的精确分子机制尚未完全阐明。现有证据表明多种蛋白质翻译后修饰在糖尿病及其并发症的发展中发挥重要作用。随着分析技术、质谱技术和定量蛋白质组学技术的发展,系统性表征生理和2型糖尿病条件下翻译后修饰的动态变化成为可能,可以为2型糖尿病及其并发症的发病机制提供分子层面的新见解。本文重点探讨了几种蛋白质翻译后修饰,包括磷酸化、酰化修饰(乙酰化、丙二酰化和琥珀酰化)、O-连接β-N-乙酰葡糖胺修饰(O-linkedβ-N-acetylglucosamine,O-GlcNAc)以及晚期糖基化终末产物(advanced glycation end products,AGEs)和酪氨酸硝基化(protein tyrosine nitration,PTN)在2型糖尿病及其并发症病理中的作用。同时综述了翻译后修饰组学技术在解析2型糖尿病的病理分子机制中的最新应用进展,为2型糖尿病发病机制研究与靶向治疗提供了新的视角。 展开更多
关键词 2型糖尿病 翻译后修饰组学 磷酸化修饰 酰化修饰 O-连接β-N-乙酰葡萄糖胺修饰 晚期糖基化终产物 酪氨酸硝化修饰
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The translation initiation factor eIF2 is phosphorylated to inhibit protein translation through reactive oxygen species under nutrient deficiencies in Arabidopsis 认领 引用
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作者 Xiaona Cui Yuanyuan Cao +4 位作者 Mengyang Lv Shuhao Zhou Meijun Chen Chengwei Li Hairong Zhang 《Stress Biology》 CAS CSCD 2025年第1期1074-1084,共11页
Nitrogen(N),phosphorus(P)or potassium(K)deficiency in plants can lead to a decrease in amino acid and protein synthesis.However,it is unknown how protein translation gets repressed during macronutrient deficiencies.Pr... Nitrogen(N),phosphorus(P)or potassium(K)deficiency in plants can lead to a decrease in amino acid and protein synthesis.However,it is unknown how protein translation gets repressed during macronutrient deficiencies.Previous research has shown that general control non-depressible 1(GCN1)cooperate with GCN2 to phosphorylate the alpha subunit of eukaryotic translation initiation factor(eIF2α).In this study,we observed phosphorylation of eIF2αunder N,P,and K deficiencies,which was found to be lost in gcn1.Mutant gcn1 displayed higher sensitivity to macronutrient deficiencies compared to the wild-type(WT).The evidence of in situ reactive oxygen species(ROS)accumulation in leaves indicated that macronutrient starvation triggers ROS production.Treatment with Dimethylthiourea(DMTU),a ROS scavenger,eliminated ROS and reversed eIF2αphosphorylation induced by nutrient deficiency.Moreover,it was discovered that protein translation was reduced under N or K deficiency in the WT but not in gcn1,whereas under P deprivation,protein translation was reduced in both the WT and gcn1.We additionally found that DMTU can partially recover translation inhibition under N or K deprivation.Taken together,it is concluded that GCN1-GCN2-eIF2αpathway is regulated by ROS and is essential for plant survival under macronutrient starvation conditions. 展开更多
关键词 Acronutrient deprivation Reactive oxygen species(ROS) General control non-depressible 1(GCN1) GCN2 eIF2αphosphorylation Protein translation
Phosphorylation of tau protein over time in rats subjected to transient brain ischemia 认领 引用 被引量：2
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作者 Bo Song Qiang Ao +6 位作者 Zhen Wang Weiqiang Liu Ying Niu Qin Shen Huancong Zuo Xiufang Zhang Yandao Gong 《Neural Regeneration Research》 SCIE CAS CSCD 2013年第34期3173-3182,共10页
Transient brain ischemia has been shown to induce hyperphosphorylation of the micro- tubule-associated protein tau. To further determine the mechanisms underlying these processes, we investigated the interaction betwe... Transient brain ischemia has been shown to induce hyperphosphorylation of the micro- tubule-associated protein tau. To further determine the mechanisms underlying these processes, we investigated the interaction between tau, glycogen synthase kinase （GSK）-313 and protein phos- phatase 2A. The results confirmed that tau protein was dephosphorylated during brain ischemia; in addition, the activity of GSK-3β was increased and the activity of protein phosphatase 2A was de- creased. After reperfusion, tau protein was hyperphosphorylated, the activity of GSK-3β was de- creased and the activity of protein phosphatase 2A remained low. Importantly, the interaction of tau with GSK-3β and protein phosphatase 2A was altered during ischemia and reperfusion. Lithium chloride could affect tau phosphorylation by regulating the interaction of tau with GSK-3β and pro- tein phosphatase 2A, and improve learning and memory ability of rats after transient brain ischemia. The present study demonstrated that it was the interaction of tau with GSK-3β and protein phos- phatase 2A, rather than their individual activities, that dominates the phosphorylation of tau in tran- sient brain ischemia. Hyperphosphorylated tau protein may play an important role in the evolution of brain injury in ischemic stroke. The neuroprotective effects of lithium chloride partly depend on the inhibition of tau phosphorylation during transient brain ischemia. 展开更多
关键词 neural regeneration brain injury brain ischemia reperfusion microtubule-associated protein tau phosphorylation glycogen synthase kinase 3[3 protein phosphatase 2A lithium chloride grants-supported paper neuroregeneration
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Serine659 in ClC-2-Target Site for Phosphorylation by MAPK 认领 引用
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作者 ZHAO Jing ZHENG Ya-juan +3 位作者 LI Gui-rong CHEN Jie YU Qian XIN Hua 《Chemical Research in Chinese Universities》 SCIE CAS CSCD 2011年第6期988-991,共4页
In order to further investigate the role of ClC-2（ClC=chloride-ion channel） played in the regulation of cell proliferation and differentiation, the capablity of ClC-2 phosphorylation catalyzed by mitogen-activated p... In order to further investigate the role of ClC-2（ClC=chloride-ion channel） played in the regulation of cell proliferation and differentiation, the capablity of ClC-2 phosphorylation catalyzed by mitogen-activated protein ki-nase（MAPK） was studied. A mutation of 659Ser to Ala（S659A） of the rabbit ClC-2 cDNA in the consensus sequence of MAPK phosphorylation was introduced by overlap extension polymerase chain reaction（PCR）. Recombinant vectors pGEX-4T-1/ClC-2-2CT and pGEX-4T-1/ClC-2CT（S659A） were constructed. They were transformed to E. coli BL21, expressed by isopropy-β-D-thiogalactoside（IPTG） induction, the recombinant proteins were subjected to purification by glutathione sepharose 4B affinity chromatography. In vitro phosphorylation of the fusion proteins catalyzed by MAPK was performed. The results show that fusion protein GST/ClC-2CT（wild type） can be phosphorylated by MAPK, and this phosphorylation can be restrained by the inhibitor p42/44MAPK, PD98095; while the phosphorylation level of fusion protein GST/ClC-2CT（S659A）（mutant） was significantly reduced. Therefore, ClC-2 can be phosphorylated by MAPK and the target site of the phosphorylation is most likely the 659Ser residue. 展开更多
关键词 ClC-2 channel Fusion protein Mitogen-activated protein kinase（MAPK） In vitro phosphorylation
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Effect of organophosphorus insecticides on phosphorylation of the M_2 muscarinic acetylcholine receptor 认领 引用
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作者 Shuyin Li Liming Zou Carry Pope 《Neural Regeneration Research》 SCIE CAS 2008年第4期406-409,共4页
BACKGROUND： Organophosphorus insecticides may promote the accumulation of acetylcholine at synapses and the neuromuscular junction by inhibiting acetylcholinesterase activity to cause disturbance of neural signal con... BACKGROUND： Organophosphorus insecticides may promote the accumulation of acetylcholine at synapses and the neuromuscular junction by inhibiting acetylcholinesterase activity to cause disturbance of neural signal conduction and induce a toxic reaction. Organophosphorus insecticides may act on M2 muscarinic acetylcholine receptors, whose combination with G proteins is regulated by phosphorylation of G protein-coupled receptor kinase 2. OBJECTIVE： To investigate the effects of organophosphorus insecticides on the phosphorylation of G protein-coupled receptor kinase 2-mediated M2 muscarinic acetylcholine receptors and to reveal other possible actions of organophosphorus insecticides. DESIGN, TIME AND SETTING： An observational study, which was performed in the Central Laboratory of Shenyang Medical College, and Department of Physiological Sciences, College of Veterinary Medicine, Oklahoma State University from June 2002 to December 2004. MATERIALS： Paraoxon, parathion, chlorpyrifos, and chlorpyrifos oxon were provided by Chem Service Company, USA, [γ -p^32] ATP and [^35S]GTP γ S by New England Nuclear Life Science Products, and recombinant β 2-adrenergic receptor membrane protein by Sigma Company, USA. METHODS： The M2 muscarinic acetylcholine receptor was extracted and purified from pig brain using affinity chromatography. Subsequently, the purified M2 muscarinic acetylcholine receptor, G protein-coupled receptor kinase 2, and [γ -p^32] ATP were incubated with different concentrations of paraoxon and chlorpyrifos oxon together. The mixture then underwent polyacrylamide gel electrophoresis, and the gel film was dried and radioactively autographed to detect phosphorylation of the M2 muscarinic acetylcholine receptor. Finally, the radio-labeled phosphorylated M2 receptor protein band was excised for counting with an isotope liquid scintillation counter. MAIN OUTCOME MEASURES： Effects of chlorpyrifos oxon, paraoxon, chlorpyrifos, and parathion in different concentrations on the phosphorylation of the M2 muscarinic acetylcholine receptor; effects of chlorpyrifos oxon on the phosphorylation of the β -adrenergic receptor. RESULTS： Chlorpyrifos oxon could completely inhibit the phosphorylation of the M2 muscarinic acetylcholine receptor, and its IC50 was 70 μ mol/L. Chlorpyrifos could also inhibit the phosphorylation of the M2 muscarinic acetylcholine receptor. However, paraoxon and parathion could not inhibit the phosphorylation of the M2 muscarinic acetylcholine receptor. Chlorpyrifos oxon in different concentrations could also not inhibit the phosphorylation of the β 2-adrenergic receptor catalyzed by G protein-coupled receptor kinase 2. CONCLUSION： Different kinds of organophosphorus insecticides have different effects on the phosphorylation of the G protein-coupled receptor kinase 2-mediated M2 muscarinic acetylcholine receptor. Organophosphorus insecticides possibly have different toxic effects. 展开更多
关键词 organophosphorus insecticide antagonists G-protein-coupled receptor kinase 2 muscarinicacetylcholine receptor M2 phosphorylation
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通窍益智颗粒通过Ca2+/CaMKⅡ信号通路改善血管性痴呆大鼠海马CA1区突触可塑性机制研究 认领 引用
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作者 陆叶 袁媛 +3 位作者 李双阳 王饶琼 唐红梅 白雪 《西部中医药》 2025年第6期22-28,共7页
目的:探讨通窍益智颗粒对血管性痴呆(vascular dementia,VaD)大鼠的相关作用机制。方法:选取92只SD大鼠,其中12只纳入假手术组,其余80只大鼠采用双侧颈总动脉永久结扎法制备VaD大鼠模型,筛选出62只造模成功的SD大鼠,按随机数字表法分为... 目的:探讨通窍益智颗粒对血管性痴呆(vascular dementia,VaD)大鼠的相关作用机制。方法:选取92只SD大鼠,其中12只纳入假手术组,其余80只大鼠采用双侧颈总动脉永久结扎法制备VaD大鼠模型,筛选出62只造模成功的SD大鼠,按随机数字表法分为模型组、美金刚组及通窍益智颗粒高、中剂量组,每组10只。假手术组、模型组予生理盐水灌胃;美金刚组大鼠以1.04 mg/(kg·d)剂量美金刚灌胃;通窍益智颗粒高、中剂量组分别以9.68、4.84 g/(kg·d)剂量通窍益智颗粒灌胃,每日灌胃2次,连续干预30天。采用Morris水迷宫方法评价大鼠行为学变化情况;透射电子显微镜观察大鼠海马CA1区神经元突触结构;流式细胞术检测大鼠海马神经元中Ca2+荧光强度;免疫荧光法测定N-甲基-D-天冬氨酸(N-methyl-D-aspartic acid receptor,NMDA)受体亚基Glu N1、Glu N2B以及α-氨基-3-羟基-5-甲基-4-异恶唑丙酸(α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid,AMPA)受体功能亚基Glu A2海马CA1区定位表达;蛋白免疫印迹法检测Glu N1、Glu N2B、Glu A2及钙调蛋白依赖性蛋白激酶(Calmodulin-dependent protein kinase,CaMKⅡ)和磷酸化钙调蛋白依赖性蛋白激酶(phosphorylation calmodulin-dependent protein kinase,p CaMKⅡ)蛋白表达情况。结果:与假手术组比较,模型组大鼠在目标象限游泳时间缩短,穿越原平台次数减少,神经元中Ca2+浓度升高,海马组织Glu N1、Glu N2B、Glu A2和p CaMKⅡ表达水平降低(P<0.05)。实验第5天模型组大鼠EL时间延长(P<0.05);与模型组比较,美金刚组及通窍益智颗粒高、中剂量组大鼠在目标象限游泳时间延长,穿越原平台次数增多,神经元中Ca2+浓度均降低,海马组织Glu N1、Glu N2B、Glu A2和p CaMKⅡ表达水平升高(P<0.05)。实验第5天美金刚组及通窍益智颗粒高、中剂量组大鼠EL时间缩短(P<0.05);与美金刚组、通窍益智颗粒高、中剂量组比较,通窍益智颗粒高剂量组各项指标改善最明显(P<0.05)。模型组大鼠海马CA1区突触间隙变模糊,突触前后膜肿胀,结构难以区分,突触活性区变小,细胞内线粒体结构破坏;通窍益智颗粒高、中剂量组和美金刚组大鼠突触结构改善,突触前后膜结构较完整,突触间隙较清晰,突触小泡增多,细胞内可见完整线粒体,以通窍益智颗粒高剂量组改善最佳。结论:基于“玄府理论”组成的通窍益智颗粒能够改善VaD模型大鼠学习记忆能力及海马CA1区神经元突触结构,其潜在机制可能与其调节Ca2+/CaMKⅡ信号通路,改善突触可塑性有关。 展开更多
关键词 血管性痴呆 玄府 钙离子 磷酸化钙调蛋白依赖性蛋白激酶 突触可塑性 通窍益智颗粒
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Regulation of Ikaros function by casein kinase 2 and protein phosphatase 1 认领 引用
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作者 Amy K Erbe Aleksandar Savic Sinisa Dovat 《World Journal of Biological Chemistry》 2011年第6期126-131,共6页
The Ikaros gene encodes a zinc finger,DNA-binding protein that regulates gene transcription and chromatin remodeling.Ikaros is a master regulator of hematopoiesis and an established tumor suppressor.Moderate alteratio... The Ikaros gene encodes a zinc finger,DNA-binding protein that regulates gene transcription and chromatin remodeling.Ikaros is a master regulator of hematopoiesis and an established tumor suppressor.Moderate alteration of Ikaros activity (e.g.haploinsufficiency) appears to be sufficient to promote malignant transformation in human hematopoietic cells.This raises questions about the mechanisms that normally regulate Ikaros function and the potential of these mechanisms to contribute to the development of leukemia.The focus of this review is the regulation of Ikaros function by phosphorylation/dephosphorylation.Site-specific phosphorylation of Ikaros by casein kinase 2 (CK2) controls Ikaros DNA-binding ability and subcellular localization.As a consequence,the ability of Ikaros to regulate cell cycle progression,chromatin remodeling,target gene expression,and thymocyte differentiation are controlled by CK2.In addition,hyperphosphorylation of Ikaros by CK2 leads to decreased Ikaros levels due to ubiquitinmediated degradation.Dephosphorylation of Ikaros by protein phosphatase 1 (PP1) acts in opposition to CK2 to increase Ikaros stability and restore Ikaros DNA binding ability and pericentromeric localization.Thus,the CK2 and PP1 pathways act in concert to regulate Ikaros activity in hematopoiesis and as a tumor suppressor.This highlights the importance of these signal transduction pathways as potential mediators of leukemogenesis via their role in regulating the activities of Ikaros. 展开更多
关键词 Ikaros Leukemia Zinc finger Transcription factor Casein kinase 2 Protein phosphatase 1 Phosphorylation
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Localization and function of a eukaryotic-initiation-factor-2-associated 67-kDa glycoprotein 认领 引用
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作者 Shiyong Wu,Edison Biotechnology Institute,Department of Chemistry and Biochemistry,Ohio University,Athens,OH 45701,United States 《World Journal of Biological Chemistry》 2010年第10期313-320,共8页
AIM: To study the localization and function of a eukaryotic initiation factor 2 (eIF2α)-associated 67-kDa glycoprotein (p67).METHODS: Immunofluorescence staining,35S-Met/Cys metabolic labeling,Western blotting analys... AIM: To study the localization and function of a eukaryotic initiation factor 2 (eIF2α)-associated 67-kDa glycoprotein (p67).METHODS: Immunofluorescence staining,35S-Met/Cys metabolic labeling,Western blotting analysis,sucrose gradient centrifugation and high speed centrifugation were used to determine the localization of proteins in transiently transfected COS-1 cells.Transient co-transfection followed by co-immunoprecipitation was used to study the interaction between p67 and double-stranded RNA (dsRNA)-dependent protein kinase (PKR).Wheat germ agglutinin agarose beads were used to absorb glycosylated proteins.In vivo 32P-labeling followed by immunoprecipitation and Western blotting were used to measure PKR autophosphorylation,eIF2α phosphorylation,and p67 expression in normal and breast cancer cells.RESULTS: The image from immunofluorescence staining showed that p67 was overexpressed in the cytosol but not in the nucleus.In a sucrose gradient,approxi-mately 30% of the overexpressed p67 was bound with ribosomes.p67 interacted with the kinase domain,butnot the dsRNA-binding domains of PKR.Only the glycosylated p67 was associated with the ribosome,and p67 did not compete with PKR for ribosome binding.In breast cancer cells,there was increased autophosphorylation of PKR but no phosphorylation of eIF2α,compared with normal breast cells.α The ratio of glycosylated/deglycosylated p67 was altered in breast cancer cells.CONCLUSION: Glycosylation of p67 is required for its ribosomal association and can potentially inhibit PKR via interaction with the kinase domain of PKR. 展开更多
关键词 Eukaryotic translation initiation factor 2 p67 Double-stranded RNA dependent protein kinase Phosphorylation Cancer
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血清Trx2、P-tau对甲状腺癌全麻术后认知损伤的预测价值 认领 引用
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作者 李聪聪 徐国亭 《华夏医学》 CAS 2025年第5期120-125,共6页
目的分析血清硫氧还蛋白2(Trx2)、磷酸化微管相关蛋白(P-tau)对甲状腺癌全麻术后认知损伤的预测价值。方法选取100例甲状腺癌患者的病例资料,依据术后是否存在认知损伤将其分为损伤组和未损伤组,分析Trx2、P-tau与甲状腺患者术后神经功... 目的分析血清硫氧还蛋白2(Trx2)、磷酸化微管相关蛋白(P-tau)对甲状腺癌全麻术后认知损伤的预测价值。方法选取100例甲状腺癌患者的病例资料,依据术后是否存在认知损伤将其分为损伤组和未损伤组,分析Trx2、P-tau与甲状腺患者术后神经功能血清指标、认知功能评分间的相关性,及对全麻术后认知损伤风险的预测效能。结果损伤组的Trx2、P-tau、神经元轴突轻链(NF-L)、神经元特异性烯醇化酶(NSE)、中枢神经特异蛋白(S100β)水平均高于未损伤组,脑源性神经营养因子(BDNF)、简易精神状态量表(MMSE)、蒙特利尔认知评估量表(MoCA)的测得值均低于未损伤组,差异均具有统计学意义(P<0.05)。Trx2、P-tau与BDNF、MMSE、MoCA均呈负相关,与NF-L、NSE、S100β均呈正相关(P<0.05)。联合检测Trx2、P-tau的灵敏度、特异度均高于Trx2、P-tau的单独检测。结论Trx2、P-tau与甲状腺癌患者全麻术后继发认知损伤呈正相关,联合检测Trx2、P-tau能实现对患者认知损伤风险的早期预测。 展开更多
关键词 甲状腺癌根治术 全身麻醉 认知损伤 硫氧还蛋白2 磷酸化微管相关蛋白
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MMP-9、p-STAT3、GLI1及SIAH2在胃腺癌组织中的表达及其临床意义分析 认领 引用
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作者 尹美辰 王晶璇 《医学理论与实践》 2025年第10期1642-1645,1661,共4页
目的:研究基质金属蛋白酶9(MMP-9)、磷酸化信号传导子机转录激活子3(p-STAT3)、胶质瘤蛋白癌基因蛋白-1(GLI1)及E3泛素蛋白连接酶2(SIAH2)在胃腺癌组织中的表达及其临床意义。方法:选取我院2019年6月—2022年6月收治的109例胃腺癌患者... 目的:研究基质金属蛋白酶9(MMP-9)、磷酸化信号传导子机转录激活子3(p-STAT3)、胶质瘤蛋白癌基因蛋白-1(GLI1)及E3泛素蛋白连接酶2(SIAH2)在胃腺癌组织中的表达及其临床意义。方法:选取我院2019年6月—2022年6月收治的109例胃腺癌患者开展研究。分别采集胃腺癌组织以及癌旁正常组织,以免疫组化染色法检测MMP-9、p-STAT3、GLI1及SIAH2表达情况并对比。分析不同病理特征胃腺癌患者MMP-9、p-STAT3、GLI1及SIAH2表达情况。结果:胃腺癌组织中MMP-9、p-STAT3、GLI1及SIAH2阳性率分别为66.06%、77.06%、74.31%、77.06%,均高于癌旁正常组织的19.27%、18.35%、21.10%、13.76%(均P0.05)。结论:MMP-9、p-STAT3、GLI1、SIAH2在胃腺癌组织中均存在异常高表达,且与病情进展密切相关。 展开更多
关键词 胃腺癌 基质金属蛋白酶9 磷酸化信号传导子机转录激活子3 胶质瘤蛋白癌基因蛋白-1 E3泛素蛋白连接酶2 临床意义
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Effects of microtubule-associated protein tau expression on neural stem cell migration after spinal cord injury 认领 引用 被引量：6
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作者 Zhi-ping Qi Guo-xiang Wang +4 位作者 Peng Xia Ting-ting Hou Hong-li Zhou Tie-jun Wang Xiao-yu Yang 《Neural Regeneration Research》 SCIE CAS CSCD 2016年第2期332-337,共6页
Our preliminary proteomics analysis suggested that expression of microtubule-associated protein tau is elevated in the spinal cord after injury.Therefore,the first aim of the present study was to examine tau expressio... Our preliminary proteomics analysis suggested that expression of microtubule-associated protein tau is elevated in the spinal cord after injury.Therefore,the first aim of the present study was to examine tau expression in the injured spinal cord.The second aim was to determine whether tau can regulate neural stem cell migration,a critical factor in the successful treatment of spinal cord injury.We established rat models of spinal cord injury and injected them with mouse hippocampal neural stem cells through the tail vein.We used immunohistochemistry to show that the expression of tau protein and the number of migrated neural stem cells were markedly increased in the injured spinal cord.Furthermore,using a Transwell assay,we showed that neural stem cell migration was not affected by an elevated tau concentration in the outer chamber,but it was decreased by changes in intracellular tau phosphorylation state.These results demonstrate that neural stem cells have targeted migration capability at the site of injury,and that although tau is not a chemokine for targeted migration of neural stem cells,intracellular tau phosphorylation/dephosphorylation can inhibit cell migration. 展开更多
关键词 nerve regeneration spinal cord injury tau protein neural stem cells transwelI chambers phosphatase 2A cell transplantation phosphorylation migration okadaic acid C2-ceramide neural regeneration
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黄连解毒汤对2型糖尿病大鼠学习记忆能力及脑内Alzheimer病样tau蛋白磷酸化途径的影响 认领 引用 被引量：16
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作者 李斌 伍文彬 +2 位作者 谢淑玲 张秀 王飞 《中华中医药杂志》 CAS CSCD 北大核心 2015年第5期1421-1425,共5页
目的:探讨黄连解毒汤对2型糖尿病(T2DM)大鼠认知功能及脑内Alzheimer病样tau蛋白磷酸化途径的影响。方法:健康雄性Wistar大鼠72只,随机分为6组,分别为空白组、模型组、罗格列酮组、黄连解毒汤低、中、高剂量组(1.5、3、6g/kg)。通过高... 目的:探讨黄连解毒汤对2型糖尿病(T2DM)大鼠认知功能及脑内Alzheimer病样tau蛋白磷酸化途径的影响。方法:健康雄性Wistar大鼠72只,随机分为6组,分别为空白组、模型组、罗格列酮组、黄连解毒汤低、中、高剂量组(1.5、3、6g/kg)。通过高糖高脂饮食及小剂量链脲佐菌素(STZ)制备T2DM模型。除空白组、模型组给予等容积的蒸馏水,其他治疗组以相应药物治疗。8周后,采用Morris水迷宫观察大鼠学习记忆能力,Western blotting法检测大鼠大脑皮层tau蛋白在不同位点的磷酸化水平。结果:黄连解毒汤可缩短T2DM模型大鼠平均逃避潜伏期(P<0.01),增加逃逸平台次数(P<0.05)、延长有效区停留时间及有效区游泳路程(P<0.05,P<0.01),有效抑制模型大鼠脑组织tau蛋白在Ser199、Ser202、Thr231、Ser396等多位点的磷酸化程度(P<0.05,P<0.01)。结论:黄连解毒汤能够有效改善T2DM模型大鼠学习和记忆能力,可在tau蛋白多位点上抑制其过度磷酸化水平。 展开更多
关键词 黄连解毒汤 2型糖尿病 阿尔茨海默病 tau蛋白磷酸化
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藤龙补中汤对大肠癌转移和整合素-FAK-ERK1/2通路蛋白表达的影响 认领 引用 被引量：9
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作者 王双双 黄晓伟 +3 位作者 李淼 陈雷 安红梅 胡兵 《中华中医药杂志》 CAS CSCD 北大核心 2020年第4期1697-1699,共3页
目的:观察藤龙补中汤对大肠癌转移和整合素-FAK-ERK1/2通路蛋白表达影响。方法:尾静脉注射人大肠癌LoVo细胞建立裸小鼠肺转移模型,随机分为对照组及中药小、大剂量组共3组,分别给予无菌水、小剂量和大剂量藤龙补中汤治疗。显微镜下观察... 目的:观察藤龙补中汤对大肠癌转移和整合素-FAK-ERK1/2通路蛋白表达影响。方法:尾静脉注射人大肠癌LoVo细胞建立裸小鼠肺转移模型,随机分为对照组及中药小、大剂量组共3组,分别给予无菌水、小剂量和大剂量藤龙补中汤治疗。显微镜下观察肺转移。免疫组织化学方法检测肺转移灶整合素αV、整合素β3、p-FAK和p-ERK1/2表达。结果:藤龙补中汤治疗后,小鼠肺转移灶显著减少(P<0.05)。藤龙补中汤同时降低LoVo大肠癌转移灶整合素αV和β3表达(P<0.01,P<0.05),抑制FAK和ERK1/2磷酸化(P<0.01)。结论:藤龙补中汤可抑制LoVo大肠癌肺转移,可能与抑制大肠癌整合素-FAK-ERK1/2信号转导相关。 展开更多
关键词 大肠癌 藤龙补中汤 转移 整合素 FAK ERK1/2 蛋白表达与磷酸化
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外源Ca2+对高温强光胁迫下小麦叶绿体D_1蛋白磷酸化及光系统Ⅱ功能的影响 认领 引用 被引量：12
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作者 李利红 马培芳 +1 位作者 杨亚军 赵会杰 《应用生态学报》 CAS 北大核心 2010年第3期683-688,共6页
用10mmol·L-1CaCl2溶液预处理灌浆期小麦叶片,以水预处理为对照,然后将预处理植株进行高温强光(35℃,1600μmol·m-2·s-1)胁迫,测定胁迫处理过程中小麦旗叶光合电子传递速率、净光合速率、叶绿素荧光参数及D1蛋白的变化,... 用10mmol·L-1CaCl2溶液预处理灌浆期小麦叶片,以水预处理为对照,然后将预处理植株进行高温强光(35℃,1600μmol·m-2·s-1)胁迫,测定胁迫处理过程中小麦旗叶光合电子传递速率、净光合速率、叶绿素荧光参数及D1蛋白的变化,以研究外源Ca2+对高温强光胁迫下小麦叶片类囊体膜D1蛋白磷酸化和PSⅡ功能的影响.结果表明:CaCl2溶液预处理使小麦叶片在高温强光逆境下PSⅡ反应中心发生可逆失活,有效抑制了高温强光下D1蛋白的净降解,保持了较高的D1蛋白磷酸化水平,暗恢复后PSⅡ反应中心活性迅速恢复,全链电子传递速率和PSⅡ电子传递速率恢复至对照水平,维持了较高的PSⅡ原初光化学效率(Fv/Fm)、实际光化学效率(ФPSⅡ)、光化学猝灭系数(qP)和净光合速率(Pn).表明外源Ca2+通过调节小麦叶绿体D1蛋白的周转,促进了PSⅡ的正常运转,减轻了高温强光胁迫对叶片光合机构的损伤. 展开更多
关键词 Ca^2+ 小麦 高温强光胁迫 D1蛋白磷酸化 光系统Ⅱ功能
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