ERK is involved in multiple cell signaling pathways through its interacting proteins. By </span><i><span style="font-size:12px;font-family:Verdana;">in</span></i> <i><s...ERK is involved in multiple cell signaling pathways through its interacting proteins. By </span><i><span style="font-size:12px;font-family:Verdana;">in</span></i> <i><span style="font-size:12px;font-family:Verdana;">silico</span></i><span style="font-size:12px;font-family:Verdana;"> analysis, earlier we have identified 22 putative ERK interacting proteins namely;ephrin type-B receptor 2 isoform 2 precursor (EPHB2), mitogen-activated protein kinase 1</span></span><span "="" style="font-size:10pt;"> </span><span "="" style="font-size:10pt;"><span style="font-size:12px;font-family:Verdana;">(MAPK1), interleukin-17 receptor D precursor (IL17RD), WD repeat domain containing 83 (WDR83), </span><span style="font-size:12px;font-family:Verdana;">tescalcin (Tesc), mitogen-activated protein kinase kinase kinase 4 (MAPP3K4),</span><span style="font-size:12px;font-family:Verdana;"> kinase suppressor of Ras2 (KSR2), mitogen-activated protein kinase kinase 6 (MAP3K6), UL16 binding protein 2 (ULBP2), UL16 binding protein 1 (ULBP1), dual specificity phosphatase 14 (DUSP14), dual specificity phosphatase 6 (DUSP6), hyaluronan-mediated motility receptor (RHAMM), kinase D interacting substrate of 220</span></span><span "="" style="font-size:10pt;"> </span><span "="" style="font-size:12px;font-family:Verdana;">kDa (KININS220), membrane-associated guanylate kinase (MAGI3), phosphoprotein enriched in astrocytes 15</span><span "="" style="font-size:10pt;"> </span><span "="" style="font-size:12px;font-family:Verdana;">(PEA15), typtophenyl-tRNA synthetase, cytoplasmic (WARS), dual specificity phosphatase 9 (DUSP9), mitogen-activated protein kinase kinase kinase 1</span><span "="" style="font-size:10pt;"> </span><span "="" style="font-size:12px;font-family:Verdana;">(MAP3K1), UL16 binding protein 3 (ULBP3), SLAM family member 7 isoform a precursor (SLAMMF7) and mitogen activated protein kinase kinase kinase 11 (MAP3K11) (</span><span "="" style="font-size:10pt;"><a href="file:///E:/%E5%B7%A5%E4%BD%9C%E8%AE%B0%E5%BD%95/2021/0225-wqs-%E5%B7%A5%E4%BD%9C%E8%AE%B0%E5%BD%95/2%E6%9C%88%20WJNS11.1%20%E6%8F%92%E9%A1%B5%E7%A0%81%20%E4%BB%98%E5%96%9C%E4%BB%81%20%EF%BC%887%EF%BC%89(1)/2%E6%9C%88%20WJNS11.1%20%E6%8F%92%E9%A1%B5%E7%A0%81%20%E4%BB%98%E5%96%9C%E4%BB%81%20%EF%BC%887%EF%BC%89/7-1390595.docx#T1"><b><span color:#943634;"="" style="font-size: 12px;font-family: Verdana;">Table 1</span></b></a></span><span "="" style="font-size:10pt;"><span style="font-size:12px;font-family:Verdana;">). However, prediction of secondary structure and domain/motif present in aforementioned ERK interacting proteins is not studied. In this paper, </span><i><span style="font-size:12px;font-family:Verdana;">in</span></i></span><i><span style="font-size:10.0pt;font-family:;" "=""> </span><span style="font-size:12px;font-family:Verdana;" "="">silico</span></i><span "="" style="font-size:12px;font-family:Verdana;"> prediction of secondary structure of ERK interacting proteins was done by SOPMA and motif/domain identification using motif search. Briefly, SOPMA predicted higher random coil and alpha helix percentage in these proteins (</span><span "="" style="font-size:10pt;"><a href="file:///E:/%E5%B7%A5%E4%BD%9C%E8%AE%B0%E5%BD%95/2021/0225-wqs-%E5%B7%A5%E4%BD%9C%E8%AE%B0%E5%BD%95/2%E6%9C%88%20WJNS11.1%20%E6%8F%92%E9%A1%B5%E7%A0%81%20%E4%BB%98%E5%96%9C%E4%BB%81%20%EF%BC%887%EF%BC%89(1)/2%E6%9C%88%20WJNS11.1%20%E6%8F%92%E9%A1%B5%E7%A0%81%20%E4%BB%98%E5%96%9C%E4%BB%81%20%EF%BC%887%EF%BC%89/7-1390595.docx#T2"><b><span color:#943634;"="" style="font-size: 12px;font-family: Verdana;">Table 2</span></b></a></span><span "="" style="font-size:12px;font-family:Verdana;">)</span><span "="" style="font-size:12px;font-family:Verdana;"> and</span><span "="" style="font-size:12px;font-family:Verdana;"> motif scan predicted serine/threonine kinases active site signature and protein kinase ATP binding region in majority of ERK interacting proteins. Moreover, few have commonly dual specificity protein phosphatase family and tyrosine specific protein phosphatase domains (</span><span "="" style="font-size:10pt;"><a href="file:///E:/%E5%B7%A5%E4%BD%9C%E8%AE%B0%E5%BD%95/2021/0225-wqs-%E5%B7%A5%E4%BD%9C%E8%AE%B0%E5%BD%95/2%E6%9C%88%20WJNS11.1%20%E6%8F%92%E9%A1%B5%E7%A0%81%20%E4%BB%98%E5%96%9C%E4%BB%81%20%EF%BC%887%EF%BC%89(1)/2%E6%9C%88%20WJNS11.1%20%E6%8F%92%E9%A1%B5%E7%A0%81%20%E4%BB%98%E5%96%9C%E4%BB%81%20%EF%BC%887%EF%BC%89/7-1390595.docx#T3"><b><span color:#943634;"="" style="font-size: 12px;font-family: Verdana;">Table 3</span></b></a></span><span "="" style="font-size:12px;font-family:Verdana;">). Such study may be helpful to design engineered molecules for regulating ERK dependent pathways in disease condition.展开更多
BACKGROUND: The increased β-arrestin-2 and its combination with G-protein-coupled receptors (GPCRs) lead to GPCRs desensitization. The latter may be responsible for decreased contractile reactivity in the mesenteric ...BACKGROUND: The increased β-arrestin-2 and its combination with G-protein-coupled receptors (GPCRs) lead to GPCRs desensitization. The latter may be responsible for decreased contractile reactivity in the mesenteric arteries of cirrhotic patients and rats. The present study is to investigate the machinery changes of α-adrenergic receptors and G proteins and their roles in the contractility of mesenteric arteries of cirrhotic patients and animal models. METHODS: Patients with cirrhosis due to hepatitis B and cirrhotic rats induced by CCl 4 were studied. Mesenteric artery contractility in response to norepinephrine was determined by a vessel perfusion system. The contractile effect of G protein-coupled receptor kinase-2 (GRK-2) inhibitor on the mesenteric artery was evaluated. The protein expression of the α 1 adrenergic receptor, G proteins, β-arrestin-2, GRK-2 as well as the activity of Rho associated coiled-coil forming protein kinase-1 (ROCK-1) were measured by Western blot. In addition, the interaction of α 1 adrenergic receptor with β-arrestin-2 was assessed by co-immunoprecipitation. RESULTS: The portal vein pressure of cirrhotic patients and rats was significantly higher than that of controls. The doseresponse curve to norepinephrine in mesenteric arteriole was shifted to the right, and EC 50 was significantly increased in cirrhotic patients and rats. There were no significant differences in the expressions of the α 1 adrenergic receptor and G proteins in the cirrhotic group compared with the controls. However, the protein expressions of GRK-2 and β-arrestin-2 were significantly elevated in cirrhotic patients and rats compared with those of the controls. The interaction of the α 1 adrenergic receptor and β-arrestin-2 was significantly aggravated. This interaction was significantly reversed by GRK-2 inhibitor. Both the protein expression and activity of ROCK-1 were significantly decreased in the mesenteric artery in patients with cirrhosis compared with those of the controls, and this phenomenon was not shown in the cirrhotic rats. Norepinephrine significantly increased the activity of ROCK-1 in normal rats but not in cirrhotic ones. Norepinephrine significantly increased ROCK-1 activity in cirrhotic rats when GRK-2 inhibitor was used. CONCLUSIONS: β-arrestin-2 expression and its interaction with GPCRs are significantly upregulated in the mesenteric arteries in patients and rats with cirrhosis. These upregulations result in GPCR desensitization, G-protein dysfunction and ROCK inhibition. These may explain the decreased contractility of the mesenteric artery in response to vasoconstrictors.展开更多
Filamentous fungi employ conserved eukaryotic signaling pathway to detect and respond to environmental signals, including the presence of the host. Genetic experiment in which a particular signaling protein is lost, o...Filamentous fungi employ conserved eukaryotic signaling pathway to detect and respond to environmental signals, including the presence of the host. Genetic experiment in which a particular signaling protein is lost, or its activity enhanced, have defined some of the function of heterotrimeric G proteins and MAP kinases in development and virulence. A hallmark of these studies is that orthologs in different species may have different functions. Antagonistic fungal-fungal interactions form the basis for biological control of plant disease. These interactions may employ novel modes of regulation by conserved signaling elements. Tag1, a G protein α subunit of Trichoderma. atroviride belonging to fungal Gi class, is involved in repression of sporulation and hyphal coiling(1). Deletion of ortholog of this gene, TgaA, in Trichoderma (Gliocladium) virens, however, did not affect sporulation and growth, yet tgaA mutants are unable to parasitize S. rolfsii sclerotia(2). Mutation of a second G α subunit gene is now under study. TmkA, a MAPK gene of T. virens, is involved in biocontrol properties and repression of conidiation(3). Using suppression-subtraction hybridization and other approaches, we are beginning to identify additional elements of the signaling cascades and their downsteam targets. The role of G protein and MAPK genes are sometimes specific to a particular host fungus or to parasitism of mycelia or sclerotia(2,3). Also of relevance to biocontrol, signal transduction pathway provide a means to alter the balance between sporulation, mycelial growth and hyphal coiling.展开更多
目的研究Rho相关卷曲螺旋蛋白激酶1(ROCK1)在动脉粥样硬化血管壁中的表达及其与基质金属蛋白酶2(MMP2)、转化生长因子1(TGF-β1)的相关性。方法选择30只载脂蛋白E基因敲除小鼠为实验组,高脂饲料喂养,另选30只C57BL/6小鼠为对照组,普通...目的研究Rho相关卷曲螺旋蛋白激酶1(ROCK1)在动脉粥样硬化血管壁中的表达及其与基质金属蛋白酶2(MMP2)、转化生长因子1(TGF-β1)的相关性。方法选择30只载脂蛋白E基因敲除小鼠为实验组,高脂饲料喂养,另选30只C57BL/6小鼠为对照组,普通饲料喂养。在喂养的第10、16、22、28及34周,取眼球血监测小鼠血脂水平;取小鼠腹主动脉作为标本,包埋切片并进行苏木精-伊红染色观察血管壁形态;应用免疫组化染色观察血管壁中ROCK1、MMP2、TGF-β1的表达;使用Image Pro Plus 6.0软件测量切片中血管壁厚度、斑块面积、血管壁中ROCK1、MMP2及TGF-β1的表达量。采用SPSS 27.0统计软件进行数据分析。采用单因素方差分析进行组间比较,两两比较采用Tukey检验。采用Pearson相关与线性回归分析ROCK1与血管壁厚度及斑块面积、MMP2、TGF-β1的关系。结果成功建立动脉粥样硬化小鼠模型。在喂养第10、16、22、28及34周,实验组血脂水平明显高于对照组,差异有统计学意义(P<0.05)。自喂养第16周起,实验组小鼠血管内均有斑块形成,随着喂养时间的延长,斑块面积和血管壁厚度不断增加,差异有统计学意义(P<0.05);血管壁内ROCK1的表达逐步增高,其表达与斑块面积及血管壁厚度呈正相关(r=0.821,0.730;P<0.05)。线性相关分析及回归分析显示,ROCK1与MMP2、TGF-β1表达均呈正相关(r=0.801,0.906;P<0.05)。结论在动脉粥样硬化血管壁中存在ROCK1蛋白的表达,且表达量随着血管壁厚度增加而增高,ROCK1的表达与MMP2、TGF-β1呈显著正相关性。鉴于ROCK1蛋白的致血管痉挛作用,提示动脉粥样硬化血管壁可能易于痉挛,具体机制需进一步研究。展开更多
文摘ERK is involved in multiple cell signaling pathways through its interacting proteins. By </span><i><span style="font-size:12px;font-family:Verdana;">in</span></i> <i><span style="font-size:12px;font-family:Verdana;">silico</span></i><span style="font-size:12px;font-family:Verdana;"> analysis, earlier we have identified 22 putative ERK interacting proteins namely;ephrin type-B receptor 2 isoform 2 precursor (EPHB2), mitogen-activated protein kinase 1</span></span><span "="" style="font-size:10pt;"> </span><span "="" style="font-size:10pt;"><span style="font-size:12px;font-family:Verdana;">(MAPK1), interleukin-17 receptor D precursor (IL17RD), WD repeat domain containing 83 (WDR83), </span><span style="font-size:12px;font-family:Verdana;">tescalcin (Tesc), mitogen-activated protein kinase kinase kinase 4 (MAPP3K4),</span><span style="font-size:12px;font-family:Verdana;"> kinase suppressor of Ras2 (KSR2), mitogen-activated protein kinase kinase 6 (MAP3K6), UL16 binding protein 2 (ULBP2), UL16 binding protein 1 (ULBP1), dual specificity phosphatase 14 (DUSP14), dual specificity phosphatase 6 (DUSP6), hyaluronan-mediated motility receptor (RHAMM), kinase D interacting substrate of 220</span></span><span "="" style="font-size:10pt;"> </span><span "="" style="font-size:12px;font-family:Verdana;">kDa (KININS220), membrane-associated guanylate kinase (MAGI3), phosphoprotein enriched in astrocytes 15</span><span "="" style="font-size:10pt;"> </span><span "="" style="font-size:12px;font-family:Verdana;">(PEA15), typtophenyl-tRNA synthetase, cytoplasmic (WARS), dual specificity phosphatase 9 (DUSP9), mitogen-activated protein kinase kinase kinase 1</span><span "="" style="font-size:10pt;"> </span><span "="" style="font-size:12px;font-family:Verdana;">(MAP3K1), UL16 binding protein 3 (ULBP3), SLAM family member 7 isoform a precursor (SLAMMF7) and mitogen activated protein kinase kinase kinase 11 (MAP3K11) (</span><span "="" style="font-size:10pt;"><a href="file:///E:/%E5%B7%A5%E4%BD%9C%E8%AE%B0%E5%BD%95/2021/0225-wqs-%E5%B7%A5%E4%BD%9C%E8%AE%B0%E5%BD%95/2%E6%9C%88%20WJNS11.1%20%E6%8F%92%E9%A1%B5%E7%A0%81%20%E4%BB%98%E5%96%9C%E4%BB%81%20%EF%BC%887%EF%BC%89(1)/2%E6%9C%88%20WJNS11.1%20%E6%8F%92%E9%A1%B5%E7%A0%81%20%E4%BB%98%E5%96%9C%E4%BB%81%20%EF%BC%887%EF%BC%89/7-1390595.docx#T1"><b><span color:#943634;"="" style="font-size: 12px;font-family: Verdana;">Table 1</span></b></a></span><span "="" style="font-size:10pt;"><span style="font-size:12px;font-family:Verdana;">). However, prediction of secondary structure and domain/motif present in aforementioned ERK interacting proteins is not studied. In this paper, </span><i><span style="font-size:12px;font-family:Verdana;">in</span></i></span><i><span style="font-size:10.0pt;font-family:;" "=""> </span><span style="font-size:12px;font-family:Verdana;" "="">silico</span></i><span "="" style="font-size:12px;font-family:Verdana;"> prediction of secondary structure of ERK interacting proteins was done by SOPMA and motif/domain identification using motif search. Briefly, SOPMA predicted higher random coil and alpha helix percentage in these proteins (</span><span "="" style="font-size:10pt;"><a href="file:///E:/%E5%B7%A5%E4%BD%9C%E8%AE%B0%E5%BD%95/2021/0225-wqs-%E5%B7%A5%E4%BD%9C%E8%AE%B0%E5%BD%95/2%E6%9C%88%20WJNS11.1%20%E6%8F%92%E9%A1%B5%E7%A0%81%20%E4%BB%98%E5%96%9C%E4%BB%81%20%EF%BC%887%EF%BC%89(1)/2%E6%9C%88%20WJNS11.1%20%E6%8F%92%E9%A1%B5%E7%A0%81%20%E4%BB%98%E5%96%9C%E4%BB%81%20%EF%BC%887%EF%BC%89/7-1390595.docx#T2"><b><span color:#943634;"="" style="font-size: 12px;font-family: Verdana;">Table 2</span></b></a></span><span "="" style="font-size:12px;font-family:Verdana;">)</span><span "="" style="font-size:12px;font-family:Verdana;"> and</span><span "="" style="font-size:12px;font-family:Verdana;"> motif scan predicted serine/threonine kinases active site signature and protein kinase ATP binding region in majority of ERK interacting proteins. Moreover, few have commonly dual specificity protein phosphatase family and tyrosine specific protein phosphatase domains (</span><span "="" style="font-size:10pt;"><a href="file:///E:/%E5%B7%A5%E4%BD%9C%E8%AE%B0%E5%BD%95/2021/0225-wqs-%E5%B7%A5%E4%BD%9C%E8%AE%B0%E5%BD%95/2%E6%9C%88%20WJNS11.1%20%E6%8F%92%E9%A1%B5%E7%A0%81%20%E4%BB%98%E5%96%9C%E4%BB%81%20%EF%BC%887%EF%BC%89(1)/2%E6%9C%88%20WJNS11.1%20%E6%8F%92%E9%A1%B5%E7%A0%81%20%E4%BB%98%E5%96%9C%E4%BB%81%20%EF%BC%887%EF%BC%89/7-1390595.docx#T3"><b><span color:#943634;"="" style="font-size: 12px;font-family: Verdana;">Table 3</span></b></a></span><span "="" style="font-size:12px;font-family:Verdana;">). Such study may be helpful to design engineered molecules for regulating ERK dependent pathways in disease condition.
基金supported by a grant from the National Natural Science Foundation of China (30972920)
文摘BACKGROUND: The increased β-arrestin-2 and its combination with G-protein-coupled receptors (GPCRs) lead to GPCRs desensitization. The latter may be responsible for decreased contractile reactivity in the mesenteric arteries of cirrhotic patients and rats. The present study is to investigate the machinery changes of α-adrenergic receptors and G proteins and their roles in the contractility of mesenteric arteries of cirrhotic patients and animal models. METHODS: Patients with cirrhosis due to hepatitis B and cirrhotic rats induced by CCl 4 were studied. Mesenteric artery contractility in response to norepinephrine was determined by a vessel perfusion system. The contractile effect of G protein-coupled receptor kinase-2 (GRK-2) inhibitor on the mesenteric artery was evaluated. The protein expression of the α 1 adrenergic receptor, G proteins, β-arrestin-2, GRK-2 as well as the activity of Rho associated coiled-coil forming protein kinase-1 (ROCK-1) were measured by Western blot. In addition, the interaction of α 1 adrenergic receptor with β-arrestin-2 was assessed by co-immunoprecipitation. RESULTS: The portal vein pressure of cirrhotic patients and rats was significantly higher than that of controls. The doseresponse curve to norepinephrine in mesenteric arteriole was shifted to the right, and EC 50 was significantly increased in cirrhotic patients and rats. There were no significant differences in the expressions of the α 1 adrenergic receptor and G proteins in the cirrhotic group compared with the controls. However, the protein expressions of GRK-2 and β-arrestin-2 were significantly elevated in cirrhotic patients and rats compared with those of the controls. The interaction of the α 1 adrenergic receptor and β-arrestin-2 was significantly aggravated. This interaction was significantly reversed by GRK-2 inhibitor. Both the protein expression and activity of ROCK-1 were significantly decreased in the mesenteric artery in patients with cirrhosis compared with those of the controls, and this phenomenon was not shown in the cirrhotic rats. Norepinephrine significantly increased the activity of ROCK-1 in normal rats but not in cirrhotic ones. Norepinephrine significantly increased ROCK-1 activity in cirrhotic rats when GRK-2 inhibitor was used. CONCLUSIONS: β-arrestin-2 expression and its interaction with GPCRs are significantly upregulated in the mesenteric arteries in patients and rats with cirrhosis. These upregulations result in GPCR desensitization, G-protein dysfunction and ROCK inhibition. These may explain the decreased contractility of the mesenteric artery in response to vasoconstrictors.
文摘Filamentous fungi employ conserved eukaryotic signaling pathway to detect and respond to environmental signals, including the presence of the host. Genetic experiment in which a particular signaling protein is lost, or its activity enhanced, have defined some of the function of heterotrimeric G proteins and MAP kinases in development and virulence. A hallmark of these studies is that orthologs in different species may have different functions. Antagonistic fungal-fungal interactions form the basis for biological control of plant disease. These interactions may employ novel modes of regulation by conserved signaling elements. Tag1, a G protein α subunit of Trichoderma. atroviride belonging to fungal Gi class, is involved in repression of sporulation and hyphal coiling(1). Deletion of ortholog of this gene, TgaA, in Trichoderma (Gliocladium) virens, however, did not affect sporulation and growth, yet tgaA mutants are unable to parasitize S. rolfsii sclerotia(2). Mutation of a second G α subunit gene is now under study. TmkA, a MAPK gene of T. virens, is involved in biocontrol properties and repression of conidiation(3). Using suppression-subtraction hybridization and other approaches, we are beginning to identify additional elements of the signaling cascades and their downsteam targets. The role of G protein and MAPK genes are sometimes specific to a particular host fungus or to parasitism of mycelia or sclerotia(2,3). Also of relevance to biocontrol, signal transduction pathway provide a means to alter the balance between sporulation, mycelial growth and hyphal coiling.
文摘目的研究Rho相关卷曲螺旋蛋白激酶1(ROCK1)在动脉粥样硬化血管壁中的表达及其与基质金属蛋白酶2(MMP2)、转化生长因子1(TGF-β1)的相关性。方法选择30只载脂蛋白E基因敲除小鼠为实验组,高脂饲料喂养,另选30只C57BL/6小鼠为对照组,普通饲料喂养。在喂养的第10、16、22、28及34周,取眼球血监测小鼠血脂水平;取小鼠腹主动脉作为标本,包埋切片并进行苏木精-伊红染色观察血管壁形态;应用免疫组化染色观察血管壁中ROCK1、MMP2、TGF-β1的表达;使用Image Pro Plus 6.0软件测量切片中血管壁厚度、斑块面积、血管壁中ROCK1、MMP2及TGF-β1的表达量。采用SPSS 27.0统计软件进行数据分析。采用单因素方差分析进行组间比较,两两比较采用Tukey检验。采用Pearson相关与线性回归分析ROCK1与血管壁厚度及斑块面积、MMP2、TGF-β1的关系。结果成功建立动脉粥样硬化小鼠模型。在喂养第10、16、22、28及34周,实验组血脂水平明显高于对照组,差异有统计学意义(P<0.05)。自喂养第16周起,实验组小鼠血管内均有斑块形成,随着喂养时间的延长,斑块面积和血管壁厚度不断增加,差异有统计学意义(P<0.05);血管壁内ROCK1的表达逐步增高,其表达与斑块面积及血管壁厚度呈正相关(r=0.821,0.730;P<0.05)。线性相关分析及回归分析显示,ROCK1与MMP2、TGF-β1表达均呈正相关(r=0.801,0.906;P<0.05)。结论在动脉粥样硬化血管壁中存在ROCK1蛋白的表达,且表达量随着血管壁厚度增加而增高,ROCK1的表达与MMP2、TGF-β1呈显著正相关性。鉴于ROCK1蛋白的致血管痉挛作用,提示动脉粥样硬化血管壁可能易于痉挛,具体机制需进一步研究。
