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1、researchpapersActaCrystallographicaSectionDStructureoftheC-terminaldomainofnsp4fromBiologicalCrystallographyfelinecoronavirusISSN0907-4449IoannisManolaridis,aJustynaA.Coronavirusesareafamilyofpositive-strandedRNAvirusesReceived15April2009aathatincludesimportantpathogensofhumansandotherAccepte
2、d14May2009Wojdyla,SantoshPanjikar,EricJ.Snijder,bAlexanderE.animals.Thelargecoronavirusgenome(2631kb)encodesGorbalenya,bHannaBerglind,c1516nonstructuralproteins(nsps)thatarederivedfromtwoPDBReference:C-terminalPa¨rNordlund,cBrunoCoutarddreplicasepolyproteinsbyautoproteolyticprocessing.Thensps
3、domainofnsp4,3gzf,r3gzfsf.aassembleintotheviralreplicationtranscriptioncomplexandandPaulA.Tucker*nsp3,nsp4andnsp6arebelievedtoanchorthisenzymecomplextomodifiedintracellularmembranes.ThelargestpartaEMBLHamburgOutstation,c/oDESY,ofthecoronavirusnsp4subunitishydrophobicandisNotkestrasse85,D-22603
4、Hamburg,Germany,predictedtobeembeddedinthemembranes.Inthisreport,bMolecularVirologyLaboratory,DepartmentofaconservedC-terminaldomain(100amino-acidresidues)MedicalMicrobiology,CenterofInfectioushasbeendelineatedthatispredictedtofacethecytoplasmandDiseases,LeidenUniversityMedicalCenter,POBox96
5、00,2300RCLeiden,hasbeenisolatedasasolubledomainusinglibrary-basedcTheNetherlands,DivisionofBiophysics,constructscreening.Aprototypicalcrystalstructureat2.8A˚DepartmentofMedicalBiochemistryandresolutionwasobtainedusingnsp4fromfelinecoronavirus.Biophysics,Scheelesva¨g2,KarolinskaInstitute,Unmod
6、ifiedandSeMet-substitutedproteinswerecrystallizeddSE-17177Stockholm,Sweden,andLaboratoireundersimilarconditions,resultingintetragonalcrystalsthatArchitectureetFonctiondesMacromole´culesBiologiques,UMR6098,AFMB–CNRS–ESIL,belongedtospacegroupP43.ThephaseproblemwasinitiallyCase925,163AvenuedeLumi
7、ny,solvedbysingleisomorphousreplacementwithanomalous13288Marseille,Francescattering(SIRAS),followedbymolecularreplacementusingaSIRAS-derivedcompositemodel.Thestructureconsistsofasingledomainwithapredominantly-helicalcontentCorrespondencee-mai
