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简介：Humanrhodopsinkinase(RK)andacarboxylterminus-truncatedmutantRKlackingthelast59aminoacids(RKC)wereexpressedinhumanembryonickidney293cellstoinvestigatetheroleofthecarboxylterminusofRKinrecognitionandphosphorylationofrhodopsin.RKC,likethewild-typeRK,wasdetectedinbothplasmamembranesandcytosolicfractions.TheCterminaltruncatedrhodopsinkinasewasunabletophosphorylatephoto-activatedrhodopsin,butpossesseskinaseactivitysimilartothewild-typeRKinphosphorylationofsmallpeptidesubstrate.ItsuggeststhatthetruncationdidnotdisturbthegrossstructuresofRKcatalyticdomain.OurresultsalsoshowthatRKCfailedtotranslocatetophoto-activatedrodoutsegments.Takentogether,ourstudydemonstratethecarboxylterminusofRKisrequiredforphosphorylationofphoto-activatedrhodopsinandstronglyindicatethatcarboxyl-terminusofRKmaybeinvolvedininteractionwithphoto-activatedrhodopsin.

简介：Ourpreviousstudiesshowedapredominanceofhighmolecularweightproteingroupintumornuclearmatrices.Contrarytonormalcells,proteinsofthisgrouparepreferentiallyphosphorylated.Phosphoproteinsofhepatomanuclearmatrixareselectivelysubjectedtorapidproteolysis.Byalkalitreatmentandamonoclonalantibodyagainstphosphotyrosylresiduethepresenceoftwohighmolecularweightbandsofphosphotyrosyl-containingproteinswasdetectedinnuclearmatricesoftumorbutnotofnormallivercells.Highmolecularweightproteingroupoftumornuclearmatricesrevealedalsoarapidturnoverandpreferentialincorporationoflabeledaminoacidsselectivelyinhibitedbychloramphenicol.

简介：增加的证据证明丝氨酸，threonine和酷氨酸残余上的蛋白质phosphorylation是在细菌的主要规章的translational以后修正。这评论在细菌的致病力集中于细菌的phosphoproteome的含意并且在phosphoproteomics和phosphorylation网络的连接加亮方法的最近的开发。在高精确性团spectrometry的最近的技术开发戏剧性地转变了proteomics并且使它可能一些彻底的地点特定的细菌的phosphoproteomes的描述。在一些细菌的phosphoproteomes的高许多酷氨酸phosphorylations在致病力建议他们的角色，特别在病原体主人相互作用的情况中；在细菌的邻蛋白质的高许多multi-phosphorylation地点是相对小的phosphorylation尺寸和蛋白质功能的精细的规定的指示物的赔偿。

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简介：ThepresentstudywasdesignedtodeterminethechangesofphosphorylationofcAMP-responseele-mentbindingprotein(CREB)inhippocampusinducedbyohmefentanylstereoisomers(F9202andF9204)inconditionedplacepreference(CPP)paradigm.TheresultsshowedthatmicereceivingF9202andF9204displayedobviousCPP.TheycouldallsignificantlystimulateCREBphosphorylationandmaintainedforalongtimewithoutaffectingtotalCREBproteinlevels.TheeffectofF9204wassimilartomorphinewhicheffectwasmorepotentandlongerthanF9202.Wealsoexaminedtheeffectsofketamine,anoncompetitiveN-mthyl-D-aspartatereceptor(NR)antagonist,onmorphine-,F9202-andF9204-inducedCPPandphos-phorylationofCREBinhippocampus.KetaminecouldsuppressnotonlytheplacepreferencebutalsothephosphorylationofCREBproducedbymorphine,F9202andF9204.ThesefindingssuggestthatalterationsinthephosphorylationofCREBberelevanttoopiatessignalingandthedevelopmentofopiatesdependence.NRantagonistsmayinterferewithopiatesdependenceandmayhavepotentialtherapeuticimplications.

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简介：Therearecurrentlynofederallyapprovedneuroprotectiveagentstotreattraumaticbraininjury.Progesterone,ahydrophobicsteroidhormone,hasbeenshowninrecentstudiestoexhibitneuroprotectiveeffectsincontrolledcorticalimpactratmodels.Aktisaproteinkinaseknowntoplayaroleincellsignalingpathwaysthatreduceedema,inflammation,apoptosis,andpromotecellgrowthinthebrain.ThisstudyaimstodetermineifprogesteronemodulatesthephosphorylationofAktviaitsthreonine308phosphorylationsite.Phosphorylationatthethreonine308siteisoneofseveralsitesresponsibleforactivatingAktandenablingtheproteinkinasetocarryoutitsneuroprotectiveeffects.ToassesstheeffectsofprogesteroneonAktphosphorylation,C57BL/6miceweretreatedwithprogesterone(8mg/kg)at1(intraperitonally),6,24,and48hours(subcutaneously)postclosed-skulltraumaticbraininjury.Thehippocampuswasharvestedat72hourspostinjuryandpreparedforwesternblotanalysis.TraumaticbraininjurycausedasignificantdecreaseinAktphosphorylationcomparedtoshamoperation.However,micetreatedwithprogesteronefollowingtraumaticbraininjuryhadanincreaseinphosphorylationofAktcomparedtotraumaticbraininjuryvehicle.Ourfindingssuggestthatprogesteroneisaviabletreatmentoptionforactivatingneuroprotectivepathwaysaftertraumaticbraininjury.

简介：到目前为止它是不清楚的是否演变氧的建筑群(OEC)的版本包括PsbO，PsbP，和PsbQ蛋白质的子单元被photosystemII(PSII)膜的phosphorylation在轻应力下面影响。在这个工作，不同phosphorylatedPSII膜从菠菜被获得。Phosphorylation部分在轻应力下面从PSII膜压制了PsbO，PsbP，和PsbQ蛋白质的版本。反应的氧种类包括superoxide阴离子，氢过氧化物和氢氧根基，在non-phosphorylated和phosphorylatedPSII膜涉及PsbO蛋白质的小部分的版本，然而并非在PsbP和PsbQ蛋白质的版本。建议的结果，PsbO，PsbP，和PsbQ蛋白质的版本部分是，这phosphorylation在PSII膜调整的所有，和在在non-phosphorylatedPSII膜的OEC子单元的版本的反应的氧种类的角色与在phosphorylatedPSII膜一样。

简介：AbstractBackground:The Nuclear Dbf2-related (NDR1) kinase is a member of the NDR/LATS family, which was a supplementary of Hippo pathway. However, whether NDR1 could inhibit glioblastoma (GBM) growth by phosphorylating Yes-associated protein (YAP) remains unknown. Meanwhile, the role of NDR1 in GBM was not clear. This study aimed to investigate the role of NDR1-YAP pathway in GBM.Methods:Bioinformation analysis and immunohistochemistry (IHC) were performed to identify the expression of NDR1 in GBM. The effect of NDR1 on cell proliferation and cell cycle was analyzed utilizing CCK-8, clone formation, immunofluorescence and flow cytometry, respectively. In addition, the xenograft tumor model was established as well. Protein interaction was examined by Co-immunoprecipitation and immunofluorescence to observe co-localization.Results:Bioinformation analysis and IHC of our patients’ tumor tissues showed that expression of NDR1 in tumor tissue was relatively lower than that in normal tissues and was positively related to a lower survival rate. NDR1 could markedly reduce the proliferation and colony formation of U87 and U251. Furthermore, the results of flow cytometry showed that NDR1 led to cell cycle arrest at the G1 phase. Tumor growth was also inhibited in xenograft nude mouse models in NDR1-overexpression group. Western blotting and immunofluorescence showed that NDR1 could integrate with and phosphorylate YAP at S127 site. Meanwhile, NDR1 could mediate apoptosis process.Conclusion:In summary, our findings point out that NDR1 functions as a tumor suppressor in GBM. NDR1 is identified as a novel regulator of YAP, which gives us an in-depth comprehension of the Hippo signaling pathway.

简介：Objective:Tostudytheeffectofnitricoxide-inducedtyrosinephosphorylationoflarge-conductancecalcium-activatedpotassium(BKCa)channelαsubunitonvascularhyporesponsivenessinrats.Methods:Atotalof46Wistarratsofeithersex,weighing250g±20g,wereusedinthisstudy.Modelsofvascularhyporesponsivenessinducedbyhemorrhagicshock(30mmHgfor2hours)invivoandbyL-arginineinvitrowereestablishedrespectively.Thevascularresponsivenessofisolatedsuperiormesentericarteriestonorepinephrinewasobserved.TyrosinephosphorylationofBKCaαsubunitwasevaluatedwithmethodsofimmunoprecipitationandWesternblotting.Results:Inthesmoothmusclecellsofthesuperiormesentericarteries,theexpressionofBKCaαsubunittyrosinephosphorylationincreasedfollowinghemorrhagicshock,andL-argininecouldinduceBKCachannelαsubunittyrosinephosphorylationinatime-anddose-dependentmanner.L-NAME(Nω-nitro-L-arginine-methyl-ester),anitricoxidesynthetaseinhibitor,couldpartlyrestorethedecreasedvasoresponsivenessofthesuperiormesentericarteriesafterhemorrhagicshockinrats.Down-regulatingtheproteintyrosinephosphorylationwithgenistein,awidely-usedspecialproteintyrosinekinaseinhibitor,couldpartlyimprovethedecreasedvasoresponsivenessofthesuperiormesentericarteriesinducedbyL-arginineinvitro,whileup-regulatingtheproteintyrosinephosphorylationwithNa3VO4,aproteintyrosinephosphataseinhibitor,couldfurtherdecreasethenitricoxide-inducedvascularhyporesponsiveness,whichcouldbepartlyamelioratedby0.1mmol/Ltetrabutylammoniumchloride(TEA),aselectiveBKCainhibitoratthisconcentration.Conclusions:NitricoxidecaninducethetyrosinephosphorylationofBKCaαsubunit,whichinfluencesthevascularhyporesponsivenessinhemorrhagicshockratsorinducedbyL-arginineinvitro.

简介：BecauseERstressisoneofthesignalingpathwaysinvolvedintheregulationofautophagy,wehypothesizedthatERstressmightalsoplayanimportantroleinradiation-inducedautophagy.SHG44andHeLacellswereirradiatedwithX-raysandcarbonionsof30and75keV/mat2Gy.TheexpressionlevelsofBip,amajorindicatorofUPR,at4and24hpost-irradiationareshowninFig.1(a)and(b)forSHG44andHeLacells,respectively.

简介：AbstractObjective:The role of Vitamin D-binding protein (DBP) in preeclampsia (PE) pathogenesis is unknown. In this study, we compared the expression of DBP in the placentas of PE patients with the placentas of normotensive pregnant women with placenta previa controls, and aimed to explore the effect of DBP on endothelial cells (ECs) and the underlying mechanism.Methods:DBP expression in placental tissues collected from PE patients and controls was evaluated by immunohistochemistry. The downregulation and upregulation of DBP expression in HTR-8/SVneo cells were examined using DBP-targeting small interfering RNA (siRNA) and DBP-expression vector, respectively. The conditioned media of these DBP-overexpressing and DBP-siRNA HTR-8/SVneo cells were collected and added to human umbilical vein EC (HUVEC) cultures. Angiogenic effects on HUVECs were assessed by tube formation assays, and the proliferation and migration of HUVECs were examined using the Real-Time Cell Analyzer. The expression of vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR)-2, as well as the phosphorylation of different residues of VEGFR-2 in HUVECs, were determined by western blotting.Results:DBP expression was significantly increased in the placental tissues collected from PE patients. The conditioned medium of DBP-overexpressing HTR-8/SVneo cells potently inhibited tube formation by HUVECs, in addition to their proliferation and migration. Furthermore, treatment of HUVECs with the conditioned medium of DBP-overexpressing HTR-8/SVneo cells decreased the phosphorylation of VEGFR-2 at tyrosine 996, whereas the treatment of these cells with the conditioned medium of DBP-siRNA HTR-8/SVneo cells increased the phosphorylation of VEGFR-2 at tyrosine 951, 996, and 1,175.Conclusions:The expression of DBP is increased in the placentas of PE patients. DBP plays potential roles in endothelial dysfunction, which contributes to PE development, by inhibiting tyrosine phosphorylation of VEGFR-2 in ECs.

简介：在生长因素刺激之上，支架蛋白质，Gab1，是酷氨酸phosphorylated并且随后适配器蛋白质，Crk，从Gab1播送信号。我们以前证明了没有细胞外的刺激，Crkoverexpression，在各种各样的人的癌症可检测，导致Gab1的酷氨酸phosphorylation。在现在的学习，内在的机制进一步被调查。CrkII的Mutational分析证明SH2领域，然而并非SH3(N)或CrkII的规章的Y221残余，为Gab1-Y307phosphorylation的正式就职是批评的。CrkII的SH2变化也减少了和Gab1的相互作用。在GST下拉试金，而Crk-SH3(N)与Gab1异种交往了，Crk-SH2跳了到野类型的Gab1，它缺乏聚类的酷氨酸区域(残余242-410)。Gab1的酷氨酸phosphorylation被表明适配器的所有Crk家庭蛋白质，然而并非另外的包含SH2导致。Src家庭kinase禁止者，PP2，废除Gab1的导致Crk的酷氨酸phosphorylations。Y307phosphorylation在缺乏Src，是，和Fyn的成纤维细胞是无法发现的，甚至在Crk的overexpression之上，而缺乏仅仅是和Fyn的房间仍然与phosphorylatedY307包含了Gab1。而且，Crk导致了Src-Y416的phosphorylation；因此，在Crk和Csk之间的相互作用被增加。Gab1-Y307F异种没能近甚至在HGF之上本地化血浆膜刺激和减少的房间移植。而且，Gab1-Y307F扰乱了Crk，FAK，和paxillin的本地化，它是焦点的粘附的典型部件。一起拿，这些结果显示Crk通过Src便于Gab1-Y307的酷氨酸phosphorylation，贡献焦点的粘附和提高的房间移植的组织，可能从而支持人的癌症开发。

简介：cAMPmediatedsignalingmayplayasuppressiveroleinimmuneresponse.WepreviouslyfoundthatthecAMP-elevators(CTxand8-Br-cAMP)inhibitedIL-12,IL-la,IL-6geneexpression,butincreasedthetranscriptionallevelsofIL-10andIL-1RainLPS-treatedmurineperitonealmacrophages.ThepresentstudyexaminedapossiblemolecularmechanisminvolvedincAMPelevators-inducedinhibitionofIL-12p40expressioninresponsetoLPS.OurdatademonstratedthatcAMPelevatorsdownregulatedIL-12p40mRNAexpressionandIL-12pT0productioninmurineperitonealmacrophages.SubsequentstudiesrevealedthatcAMP-elevatorsblockedphosphorylationofp38MAPK,butdidnotaffecttheactivityofNF-κBbindingtoIL-12promoter(-136/-112).ThisisthefirstreportthatcAMPelevatorsinhibitLPS-inducedIL-12productionbyamechanismthatisassociated,atleastinpart,withp38-dependentinhibitionbycAMPsignalingpathways.