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.
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.
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.
简介:AbstractSwine acute diarrhea syndrome coronavirus (SADS-CoV) is a recently discovered coronavirus that causes severe and acute diarrhea and rapid weight loss in piglets. SADS-CoV was reported to be capable of infecting cell lines derived from diverse species, including bats, mice, hamsters, rats, chickens, pigs, nonhuman primates, and humans, implying its high risk of cross-species infection. However, its receptor is still unknown. In this study, the receptor-binding domain of the SADS-CoV spike (S) protein was purified and then subjected to affinity purification (AP)-coupled mass spectrometry (MS)-based proteomic analysis to identify the interactors of the SADS-CoV S protein. Forty-three host proteins were identified, and a Gene Ontology analysis indicated that these interactors can be grouped into categories such as "cell-cell adhesion" , "translation" "viral transcription" , suggesting that these processes may participate in the SADS-CoV life cycles. RNA interference-based screening of these interactors indicated that PPIB and vimentin can affect SADS-CoV replication. Our study provides an overarching view into the host interactome of the SADS-CoV S protein and highlights potential targets for the development of therapeutics against SADS-CoV.
简介:AbstractNonalcoholic fatty liver disease (NAFLD) is becoming increasingly common as the global economy grows and living standards improve. Timely and effective preventions and treatments for NAFLD are urgently needed. Retinol-binding protein-4 (RBP4), the protein that transports retinol through the circulation, was found to be positively related to diabetes, obesity, cardiovascular disease, and other metabolic diseases. Observational studies on the association between serum RBP4 level and the prevalence of NAFLD found contradictory results. Some of the underlying mechanisms responsible for this association have been revealed, and the possible clinical implications of treating NAFLD by targeting RBP4 have been demonstrated. Future studies should focus on the predictive value of RBP4 on NAFLD development and its potential as a therapeutic target in NAFLD.
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简介:SolublemalewormantigenofSchistosomajaponicurn(Sj)wasinvestigatedfordevelopmentofnewvaccinecandidate.SDS--PAGEandWesternblottingwereperformedtocomparethedifferencebetweensolubleantigensfromwormsofdifferentsex.MicevaccinationwiththetestingpurifiedproteinwasfollowedbySjcereariaechallengetodetecttheprotectiveeffectagainstSj.Sixteenbandswereseenforthesolublemalewormantigenand12forthefemaleworm.Inaddition,adistinctbandof44.6kDafromthemalewormantigenwasobserved,anditsantigenicitywasdemonstratedbyWesternblotting.This44.6kDaproteincouldinducesignificantwormandeggreductionrateinmice(39.31%,41.98%,P<0.001).Inthisstudya44.6kDaproteinwasisolatedandpartiallycharacterized.Itsantigenicity,immunogenicityandthepartialimmuneprotectionsuggestitspotentialvaccinecandidteagainstSj.