Molecular Dynamics of Mesophilic-Like Mutants of a Cold-Adapted Enzyme Insights into Distal Effects Induced by the Mutations 英文参考文献.docVIP

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Molecular Dynamics of Mesophilic-Like Mutants of a Cold-Adapted Enzyme Insights into Distal Effects Induced by the Mutations 英文参考文献.doc

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Molecular Dynamics of Mesophilic-Like Mutants of a Cold-Adapted Enzyme Insights into Distal Effects Induced by the Mutations 英文参考文献

MolecularDynamicsofMesophilic-LikeMutantsofa Cold-AdaptedEnzyme:InsightsintoDistalEffects InducedbytheMutations ElenaPapaleo*,MarcoPasi¤,MatteoTiberti,LucaDeGioia DepartmentofBiotechnologyandBiosciences,UniversityofMilano-Bicocca,Milan,Italy Abstract Networks and clusters of intramolecular interactions, as well as their ‘‘communication’’ across the three-dimensional architecturehaveaprominentroleindeterminingproteinstabilityandfunction.Specialattentionhasbeendedicatedto theirroleinthermaladaptation.Inthepresentcontribution,sevenpreviouslyexperimentallycharacterizedmutantsofa cold-adapted a-amylase, featuring mesophilic-like behavior, have been investigated by multiple molecular dynamics simulations, essential dynamics and analyses of correlated motions and electrostatic interactions. Our data elucidate the molecularmechanismsunderlyingtheabilityofsingleandmultiplemutationstogloballymodulatedynamicpropertiesof thecold-adapteda-amylase,includingbothlocalandcomplexunpredictabledistaleffects.Ourinvestigationalsoshows,in agreementwiththeexperimentaldata,thattheconversionofthecold-adaptedenzymeinawarm-adaptedvariantcannot becompletelyachievedbytheintroductionoffewmutations,alsoprovidingtherationalebehindtheseeffects.Moreover, pivotalresidues,whicharelikelytomediatetheeffectsinducedbythemutations,havebeenidentifiedfromouranalyses,as wellasagroupofsuitablecandidatesforproteinengineering.Infact,asubsetofresidueshereidentified(asanisoleucine, or networks of mesophilic-like salt bridges in the proximity of the catalytic site) should be considered, in experimental studies,togetamoreefficientmodificationofthefeaturesofthecold-adaptedenzyme. Citation:PapaleoE,PasiM,TibertiM,DeGioiaL(2011)MolecularDynamicsofMesophilic-LikeMutantsofaCold-AdaptedEnzyme:InsightsintoDistalEffects InducedbytheMutations.PLoSONE6(9):e24214.doi:10.1371/journal.pone.0024214 Editor:FrancaFraternali,King9sCollegeLondon,UnitedKingdom ReceivedApril25,2011;AcceptedAugust2,2011;PublishedSeptember7,2011 Copyright