Genetic PEGylation 英文参考文献.docVIP

GeneticPEGylation SeiichiTada1,TakashiAndou1,2,TakehiroSuzuki3,NaoshiDohmae3,EiryKobatake2,YoshihiroIto1,2* 1NanoMedicalEngineeringLaboratory,RIKENAdvancedScienceInstitute,Wako,Saitama,Japan,2GraduateSchoolofBioscienceandBiotechnology,TokyoInstituteof Technology,Yokohama,Kanagawa,Japan,3BiomolecularCharacterizationTeam,ChemicalBiologyCoreFacility,ChemicalBiologyDepartment,RIKENAdvancedScience Institute,Wako,Saitama,Japan Abstract Polyethyleneglycol(PEG)wasgeneticallyincorporatedintoapolypeptide.Stop-anticodon-containingtRNAswereacylated withPEG-containingaminoacidsandwerethentranslatedintopolypeptidescorrespondingtoDNAsequencescontaining thestopcodons.ThemolecularweightsofthePEGusedwere170,500,700,1000,and2000Da,andthetranslationwas confirmedbymassspectrometry.ThePEGincorporationratiodecreasedasthemolecularweightofPEGincreased,andPEG with a molecular weight of 1000Da was only slightly incorporated. Although improvement is required to increase the efficiencyoftheprocess,thisstudydemonstratesthepossibilityofgeneticPEGylation. Citation:TadaS,AndouT,SuzukiT,DohmaeN,KobatakeE,etal.(2012)GeneticPEGylation.PLoSONE7(11):e49235.doi:10.1371/journal.pone.0049235 Editor:NediljkoBudisa,BerlinInstituteofTechnology,Germany ReceivedMarch19,2012;AcceptedOctober8,2012;PublishedNovember8,2012 Copyright: ? 2012 Tada etal. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:Theseauthorshavenosupportorfundingtoreport. CompetingInterests:Theauthorshavedeclaredthatnocompetinginterestsexist. *E-mail:y-ito@riken.jp Introduction complementary tRNA. Deiters etal. [44] made 20 different versionsofhumangrowthfactor,eachofwhichhadanonnatural aminoacid,p-acetylphenylalanine,insertedatadifferentsiteusing the misacylated tRNA method. They linked a single PEG molecule to each keto group as a posttranslational modification.