简介：无

简介：在精子房间的膜修正在精子capacitation代表关键步；然而，这些修正的分子的基础充分没被理解。Ezrin是ezrin/radixin/merlin家庭的学习最好的成员。作为在外皮的细胞骨架和血浆膜蛋白质之间的cross-linker，ezrin贡献膜表面结构改变。而且，激活ezrin和Rho分离禁止者，RhoGDI，通过Rho激活支持外皮的cytoskeleton-polymerized肌动朊的形成。因此，ezrin，肌动朊，RhoGDI，Rho和血浆膜蛋白质在vivo形成一个复杂网络，它贡献膜表面的结构的集会。以前，我们证明ezrin和RhoGDI1在人的睾丸被表示。因此，我们寻求了决定ezrin鈥揜hoGDI1鈥揳c听鈥搈embrane蛋白质网络是否在人的精子capacitation有一个角色。我们由西方的污点的结果显示ezrin被threonine567残余的phosphorylation在capacitation期间激活。Co-immunoprecipitation研究表明在精子capacitation期间，在ezrin和RhoGDI1之间的相互作用增加，并且二维的电气泳动胶化的phosphostaining证明RhoGDI1是phosphorylated，建议RhoGDI1从RhoA分裂并且导致精子头上的肌动朊聚合。我们推测那激活的ezrin在capacitation以后与polymerized肌动朊和glycosylated膜蛋白质cd44交往。用ezrin特定或肌动朊特定的monoclonal抗体堵住精子capacitation减少他们的acrosome反应(AR)率，但是没独自在AR上有效果。一起拿，我们的结果证明由ezrin，RhoGDI1，RhoA，F肌动朊和膜蛋白质组成的一个网络工作影响在人的精子capacitation期间发生在精子头的膜上的修正。

简介：房间有大量的控制维持他们的完整并且阻止随机切换到另外一个一个生物状态。RafKinase禁止的蛋白质(RKIP)，phosphatidylethanolamine绑定蛋白质(PEBP)的一个成员家庭，表明工作维持“yinyang”或生物系统的平衡的串联的调节的人的一个新类是代表性的。RKIP禁止地图kinase(Raf-MEK-ERK)，表明串联的G联合蛋白质的受体(GPCR)kinase和NFkappaB。因为RKIP指向依赖于它磷酸化的状态的不同家族ases，RKIP也行动集成多重环境刺激开始的串音。RKIP的损失或弄空导致正常细胞的壅滞和罐头的混乱导致象癌症那样的chromosomal畸形和疾病状态。因为RKIP和PEBP家庭以前被考察了，这分析的目标是提供更改并且加亮一些RKIP的唯一的特征在细胞的发信号的过程的规定使它成为一个批评播放器。

简介：Theinteractionofbromothymolblue(BB)withhumanserumalbumin(HSA)wasstudiedbyelectrochemicaltechniquesandasensitivemethodforproteinsassaywasdeveloped.WhenBBinteractedwithHSA,thevoltammetricpeakcurrentvalueofBBdecreasedlinearlywiththeconcentrationofHSAinarangeof1.0―40.0mg/L,andthepeakpotentialshiftednegatively.Basedontheresults,asensitiveassaymethodforproteins,suchasHSA,bovineserumalbumin(BSA),andeggalbuminetc.wasestablished.ThismethodwasfurtherappliedtodeterminingtheHSAinhealthyhumanbloodsamples,andtheresultsarenotsignificantlydifferentfromthoseobtainedbytheclassicCoomassieBrilliantBlueG-250spectrophotometicmethod.Thedetectingconditionsofthismethodwereoptimizedandtheinteractionmechanismwasdiscussed.Theresultsshowthattheelectrochemicalparameters(formalpotentialE0,standardrateconstantoftheelectrodereactionks,parameterofkineticnα)ofBBhavenoobviouschangesbeforeandaftertheinteraction,whichindicatethatBBcaninteractwithHSA,forminganelectrochemicalnon-activecomplex.Theequilibriumconstant(βs)andthebindingratio(m)forthiscomplexwerecalculated.Themis4andβsis1.41×1019.Thismethodisfast,simple,highlysensitive,andhasgoodselectivity,whichcanbeusedinclinicalmeasurements.

简介：TheCoronaviridaefamilyischaracterizedbyanucleocapsidthatiscomposedofthegenomeRNAmoleculeincombinationwiththenucleoprotein(Nprotein)withinavirion.ThemoststrikingphysiochemicalfeatureoftheNproteinofSARS-CoVisthatitisatypicalbasicproteinwithahighpredictedpIandhighhydrophilicity,whichisconsistentwithitsfunctionofbindingtotheribophosphatebackboneoftheRNAmolecule.ThepredictedhighextentofphosphorylationoftheNproteinonmultiplecandidatephosphorylationsitesdemonstratesthatitwouldberelatedtoimportantfunctions,suchasRNA-bindingandlocalizationtothenucleolusofhostcells.SubsequentstudyshowsthatthereisanSR-richregionintheNproteinandthisregionmightbeinvolvedintheprotein-proteininteraction.TheabundantantigenicsitespredictedintheNprotein,aswellasexperimentalevidencewithsynthesizedpolypeptides,indicatethattheNproteinisoneofthemajorantigensoftheSARS-CoV.Comparedwithotherviralstructuralproteins,thelowvariationrateoftheNproteinwithregardstoitssizesuggestsitsimportancetothesurvivalofthevirus.

简介：唯一的肽(杯)的一个概念被建议并且实现了从双人脚踏车团spectrometry(MS/MS)识别整个房间的蛋白质离子系列。唯一的肽被定义为肽，不管它的长度，那仅仅在兴趣的proteome的一蛋白质存在，尽管这肽可以在一样的蛋白质非常出现一次。集成杯，一个二拍子的圆舞整个房间的蛋白质鉴定策略被开发进一步增加识别蛋白质的信心。包括吉祥物和SEQUEST包含Saccharomycescerevisiae和蛋白质鉴定工具的40,243个MS/MS离子系列的数据集被用来说明建议概念和策略。没有实现杯，SEQUEST识别的蛋白质是吉祥物识别的那些的2.26褶层。当杯被使用时，忍受唯一的肽的蛋白质由SEQUEST鉴别那些的3.89褶层被吉祥物识别。由跨comparing识别蛋白质的二个集合，仅仅从杯导出的89普通蛋白质被发现。在识别蛋白质之间的关键差异从每个蛋白质鉴定工具采用的filterng标准被结果。根据杯和蛋白质鉴定工具认出的蛋白质的公共分类的肽的起源，所有识别蛋白质是跨compared，导致拥有分配信心的不同层次的四组蛋白质。

简介：Hausdorffdistancebetweentwocompactsets,definedasthemaximumdistancefromapointofonesettoanotherset,hasmanyapplicationincomputerscience.Itisagoodmeasureforthesimilarityoftwosets.ThispaperprovesthattheshapedistancebetweentwocompactsetsinR~ndefinedbyminimumHausdorffdistanceunderrigidmotionsisadistance.Theauthorsintroducesimilaritycomparisonproblemsinproteinscience,andproposethatthismeasuremayhavegoodapplicationtocomparisonofproteinstructureaswell.Forcalculationofthisdistance,theauthorsgiveonedimensionalformulasforproblems(2,n),(3,3),and(3,4).Theseformulascanreducetimeneededforsolvingtheseproblems.Theauthorsdidsomenumericalexperimentsfor(2,n).Onthesesetsofdata,thisformulacanreducetimeneededtoonefifteenthofthebestalgorithmsknownonaverage.Asnincreases,itwouldsavemoretime.

简介：RecognitionofDNAdamageisacriticalstepforDNAdamage-mediatedcellularresponse.XPCisanimportantDNAdamagerecognitionproteininvolvedinnucleotideexcisionrepair(NER).WehavestudiedtheXPCproteinincisplatinDNAdamagingtreatment-mediatedcellularresponse.ComparisonofthemicroarraydatafrombothnormalandXPCdefectivehumanfibroblastsidentified861XPC-responsivegenesinthecisplatintreatment(withminimumfoldchange≥1.5).Thecellcycleandcellproliferation-relatedgenesarethemostaffectedgenesbytheXPCdefectinthetreatment.Manyothercellularfunctiongenes,especiallytheDNArepairandsignaltransduction-relatedgenes,werealsoaffectedbytheXPCdefectinthetreatment.Tovalidatethemicroarraydata,thetranscriptionlevelsofsomemicroarray-identifiedgeneswerealsodeterminedbyanRT-PCRbasedrealtimePCRassay.TherealtimePCRresultsareconsistentwiththemicroarraydataformostofthetestedgenes,indicatingthereliabilityofthemicroarraydata.Tofurthervalidatethemicroarraydata,thecisplatintreatment-mediatedcaspase-3activationwasalsodetermined.TheWesternblothybridizationresultsindicatethattheXPCdefectgreatlyattenuatesthecisplatintreatment-mediatedCaspase-3activation.Weelucidatedtheroleofp53proteinintheXPCproteinDNAdamagerecognition-mediatedsignalingprocess.TheXPCdefectreducesthecisplatintreatment-mediatedp53response.TheseresultssuggestthattheXPCproteinplaysanimportantroleinthecisplatintreatment-mediatedcellularresponse.Itmayalsosuggestapossiblemechanismofcancercelldrugresistance.

简介：在公孙树bilobaseeds的内乳的蛋白质多肽的动态变化被SDS页和二维的胶化电气泳动(2-DE)在种子萌芽期间学习。结果证明在公孙树biloba的内乳的80种蛋白质点是在2-DE光谱的clearobserved。蛋白质分子的重量在26的范围—52kD，和theirisoelectric点在5.8-7.8的范围。在种子萌芽期间，13类型ofproteins被降级，并且13种蛋白质被综合；7种蛋白质withdifferent分子的重量和35kD/pI6.8的等电位的点，31kD/pI6.8，29kD/pI6.8,33kD/pI6.6，33kD/pI6.4，34kD/pI7.7和31kD/pI7.7首先作为植物的storageproteins(VSP)被识别。

简介：WestudiedstructuralandimmunologicalpropertiesoftheSARS-CoVM(mem-brane)protein,basedoncomparativeanalysesofsequencefeatures,phylogeneticinvestigation,andexperimentalresults.TheMproteinispredictedtocontainatriple-spanningtransmembrane(TM)region,asingleN-glycosylationsitenearitsN-terminusthatisintheexteriorofthevirion,andalongC-terminalregionintheinterior.TheMproteinharborsahighersubstitutionrate(0.6%correlatedtoitssize)amongviralopenreadingframes(ORFs)frompublisheddata.ThefoursubstitutionsdetectedintheMprotein,whichcausenon-synonymouschanges,canbeclassifiedintothreetypes.OneofthemresultsinchangesofpI(isoelectricpoint)andcharge,affectingantigenicity.ThesecondchangeshydrophobicityoftheTMregion,andthethirdonerelatestohydrophilicityoftheinteriorstructure.PhylogenetictreebuildingbasedonthevariationsoftheMproteinappearstosupportthenon-humanoriginofSARS-CoV.Toinvestigateitsimmunogenicity,wesynthesizedeightoligopeptidescovering69.2%oftheentireORFandscreenedthembyusingELISA(enzyme-linkedimmunosorbentassay)withserafromSARSpatients.Theresultsconfirmedourpredictionsonantigenicsites.

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简介：ThemuscleproteinmyosinbindingproteinC(MyBPC)isalargemulti-domainproteinwhoseroleinthesarcomereiscomplexandnotyetfullyunderstood.MutationsinMyBPCarestronglyassociatedwiththeheartdiseasefamilialhypertrophiccardiomyopathy(FHC)andtheseexperimentsofnaturehaveprovidedsomeinsightintotheintricateworkingsofthisproteinintheheart.WhilesomeregionsoftheMyBPCmoleculehavebeenassignedafunctionintheregulationofmusclecontraction,theinteractionofotherregionswithvariouspartsofthemyosinmoleculeandthesarcomericproteins,actinandtitin,remainobscure.Inadditicn,severalintra-domaininteractionsbetweenadjacentMyBPCmoleculeshavebeenidentified.Althoughthebasicstructureofthemolecule(aseriesofimmunoglobulinandfibronectindomains)hasbeenelucidated,theassemblyofMyBPCinthesarcomereisatopicfordebate.ByanalysingtheMyBPCsequencewithrespecttoFHC-causingmutationsitispossibletoidentifyindividualresiduesorregionsofeachdomainthatmaybeimportanteitherforbindingorregulation.Thisreviewlooksatthecurrentliterature,inconcertwithalignmentsandthestructuralmodelsofMyBPC,inanattempttounderstandhowFHCmutationsmayleadtothediseasestate.

简介：Sumoylationisanimportantproteinmodificationdiscoveredrecently.SUMO(smallubiquitin-relatedmodifier)pathwayregulatestheproteinstabilityandtranscriptionalactivitywitha12-kDasmallmolecularprotein,SUMO,ligatedtothetargetprotein.ThepurificationofSUMOproteinsisakeysteptorevealtheirfunction.ThepurposeofthisstudywastoconstructtherecombinantSUMO1geneclonedtoapGEX-4T-1vectortoexpressandpurifytheSUMO1-GSTfusionproteininEscherichiacoli.First,thefulllengthDNAsequenceofSUMO1genewasamplifiedbyPCRandwasligatedtopMD18-Tvector.ThentheSUMO1genewassubclonedtopGEX-4T-1prokaryoticexpressionvectorbetweenBamHIandXhoIsites,andtransformedinEscherichiacoliDH5αcells.Therightcolonieswereidentifiedbyrestrictiveenzymedigestionandsequencing.ThecorrectrebombinantplasmidofpGEX-4T-1-SUMO1wastransformedinEscherichiacoliBL21cellsandtheninducedbyIPTG(isopropyl-β-D-1-thiogalacto-pyranoside)toexpresstheSUMO1-GSTfusionprotein.ThehighlypurifiedSUMO1-GST(glutathioneS-transferase)fusionproteinwasobtainedbyaffinitychromatography.Finally,thepropertiesofSUMO1-GSTfusionproteinwereconfirmedbyCoomassiebrilliantbluestrainandWesternblotanalysis.TherecombinantplasmidofpGEX-4T-1-SUMO1wassuccessfullyconstructed,andSUMO1-GSTfusionproteinsweresuccessfullyexpressed.

简介：Inthefaceoftheworldwidethreatofsevereacuterespiratorysyndrome(SARS)tohumanlife,someofthemosturgentchallengesaretodevelopfastandaccurateanalyticalmethodsforearlydiagnosisofthisdiseaseaswellastocreateasafeanti-viralvaccineforprevention.Totheseends,weinvestigatedtheantigenicityofthespikeprotein(Sprotein),amajorstructuralproteinintheSARS-coronavirus(SARS-CoV).BaseduponthetheoreticalanalysisforhydrophobicityoftheSprotein,18peptidesweresynthesized.UsingEnzyme-LinkedImmunosorbentAssay(ELISA),thesepeptideswerescreenedintheserafromSARSpatients.Accordingtotheseresults,twofragmentsoftheSgenewereamplifiedbyPCRandclonedintopET-32a.BothSfragmentswereexpressedintheBL-21strainandfurtherpurifiedwithanaffinitychromatography.TheserecombinantSfragmentswereconfirmedtohavepositivecross-reactionswithSARSsera,eitherbyWesternblotorbyELISA.OurresultsdemonstratedthatthepotentialepitoperegionswerelocatedatCodons469-882intheSprotein,andoneepitopesitewaslocatedatCodons599-620.IdentificationofantigenicregionsintheSARS-CoVSproteinmaybeimportantforthefunctionalstudiesofthisvirusorthedevelopmentofclinicaldiagnosis.

简介：Alaskapollockisanimportantproteinsourcewhichisextensivelyusedinthefoodindustry.Pollockproteinisolates(PPI)withsignificantlyenrichedproteincontentscouldbepreparedusingisoelectricsolubilization/precipitation(ISP)processing;however,thefunctionalpropertiesofthisprocessislimitedbythelargeamountofwater-insolubleproteins.Inthisstudy,weinvestigatedtheinfluenceofhighhydrostaticpressure(HHP)treatmentonthesolubilityandstructuralchangesofPPI.PPIobtainedusingISPistreatedwithhydrostaticpressuresof200,300,400,and500MPaforupto15min,andtheHHP-treatedsampleswereobservedtoexhibitsignificantlyimprovedsolubilities.FurtherbiochemicalassaysrevealthatthecontinuousHHPtreatmentsreducethecontentsoffreesulfhydrylgroupsandpromotetheformationofmacromoleculeswithbetterwatersolubilities,whichmayinducethesolubilityimprovementsoftheHHP-treatedPPI.OurresultsindicatethatHHPcanbeutilizedtoeffectivelypreparehighlywater-solubleAlaskapollockproteininfoodprocessing.

简介：Abranchandboundalgorithmisproposedforthetwo-dimensionalproteinfoldingproblemintheHPlatticemodel.Inthisalgorithm,thebenefitofeachpossiblelocationofhydrophobicmonomersisevaluatedandonlypromisingnodesarekeptforfurtherbranchingateachlevel.Theproposedalgorithmiscomparedwithotherwell-knownmethodsfor10benchmarksequenceswithlengthsrangingfrom20to100monomers.Theresultsindicatethatourmethodisaveryefficientandpromisingtoolfortheproteinfoldingproblem.

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简介：AbstractBackground:Mounting evidence, consistent with our previous study, showed that γ-aminobutyric acid type A receptor (GABAAR) played an indispensable role in airway inflammation and mucus hypersecretion in asthma. Monocyte chemotactic protein-inducing protein 1 (MCPIP1) was a key negative regulator of inflammation. Recent studies showed that inflammation was largely suppressed by enhanced MCPIP1 expression in many inflammatory diseases. However, the role and potential mechanism of MCPIP1 in airway inflammation and mucus hypersecretion in asthma were still not well studied. This study was to explore the role of MCPIP1 in asthmatic airway inflammation and mucus hypersecretion in both mice and BEAS-2B cells, and its potential mechanism.Methods:In vivo, mice were sensitized and challenged by ovalbumin (OVA) to induce asthma. Airway inflammation and mucus secretion were analyzed. In vitro, BEAS-2B cells were chosen. Interleukin (IL)-13 was used to stimulate inflammation and mucus hypersecretion in cells. MCPIP1 Lentiviral vector (LA-MCPIP1) and plasmid-MCPIP1 were used to up-regulate MCPIP1 in lung and cells, respectively. MCP-1, thymic stromal lymphopoietin (TSLP), mucin 5AC (MUC5AC), MCPIP1, and GABAARβ2 expressions were measured in both lung and BEAS-2B cells. Immunofluorescence staining was performed to observe the expression of GABAARβ2 in cells.Results:MCPIP1 was up-regulated by LA-MCPIP1 (P < 0.001) and plasmid-MCPIP1 (P < 0.001) in lung and cells, respectively. OVA-induced airway inflammation and mucus hypersecretion, OVA-enhanced MCP-1, TSLP, MUC5AC, and GABAARβ2 expressions, and OVA-reduced MCPIP1 were significantly blunted by LA-MCPIP1 in mice (all P < 0.001). IL-13-enhanced MCP-1, TSLP, MUC5AC, and GABAARβ2 expressions, and IL-13-reduced MCPIP1 were markedly abrogated by plasmid-MCPIP1 in BEAS-2B cells (all P < 0.001).Conclusion:The results of this study suggested that OVA and IL-13-induced airway inflammation and mucus hypersecretion were negatively regulated by MCPIP1 in both lung and BEAS-2B cells, involving GABAAR signaling pathway.

简介：Preparationandcharacterizationofthehapten-proteinconjugatesarefundamentaltodevelopingenvironmentalimmunoassays.Asahapten,1-pyrenebutyricacid(PBA)wasconjugatedtothecarrierproteinofbovineserumalbumin(BSA)orovalbumin(OVA)byactiveestermethod.Infraredspectra(IR)showedthatPBA-BSAandPBA-OVAconjugatesweresuccessfullyprepared.Thenumberofthehaptensconjugatedtothecarrierproteinwasdeterminedbyultravioletspectra(UV)andmatrix-assistedlaserdesorptionionizationtime-of-flightmassspectrometry(MALDI-TOF-MS).ThecalculatedaveragebindingratiosofPBA/BSAandPBA/OVAwere18:1and10:1byUV,and31:1and22:1byMALDI-TOF-MS,respectively.Althoughtherewasadiscrepancybetweentheresultsdeterminedbythetwomethods,bothofthemwereusefulforthecharacterizationofthehapten-proteinconjugates.TheantibodywasproducedagainsttheantigenofPBA-BSA,andtheaffinitywastestedbythedoubleagardiffusionmethod.Theconjugatesandtheantibodycouldbeusedfordevelopingasensitiveandselectiveimmunoassayofpolycyclicaromatichydrocarbons(PAHs).

简介：ErbB2,amemberofthereceptortyrosinekinasefamily,isfrequentlyover-expressedinbreastcancer.ProteolysisoftheextracellulardomainofErbB2resultsinconstitutiveactivationofErbB2kinase.RecentstudyreportedthatErbB2isfoundinthenucleus.Here,weshowedthatErbB2isimportedintothenucleusthroughanuclearlocalizationsignal(NLS)-mediatedmechanism.TheNLSsequenceKRRQQKIRKYTMRR(aa655-668)containsthreeclustersofbasicaminoacidsanditissufficienttotargetGFPintothenucleus.However,mutationinanybasicaminoacidclusterofthisNLSsequencesignificantlyaffectsitsnuclearlocalization.Furthermore,itwasfoundthatthisNLSisessentialforthenuclearlocalizationofErbB2sincetheintracellulardomainofErb2lackingNLScompletelyabrogatesitsnucleartranslocation.Takentogether,ourstudyidentifiedanovelnuclearlocalizationsignalandrevealsanovelmechanismunderlyingErbB2nucleartraffickingandlocalization.