Rational Engineering of Enzyme Allosteric Regulation through Sequence Evolution Analysis 英文参考文献.docVIP
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Rational Engineering of Enzyme Allosteric Regulation through Sequence Evolution Analysis 英文参考文献
RationalEngineeringofEnzymeAllostericRegulation
throughSequenceEvolutionAnalysis
Jae-SeongYang1.,SangWooSeo2.,SunghoJang2,GyooYeolJung1,2*,SangukKim1,3,4
*
1School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Gyeongbuk, Korea, 2Department of Chemical
Engineering, Pohang University of Science and Technology, Pohang, Gyeongbuk, Korea, 3Division of Molecular and Life Science, Pohang University of Science and
Technology,Pohang,Gyeongbuk,Korea,4DivisionofITConvergenceEngineering,PohangUniversityofScienceandTechnology,Pohang,Gyeongbuk,Korea
Abstract
Control of enzyme allosteric regulation is required to drive metabolic flux toward desired levels. Although the three-
dimensional (3D) structures of many enzyme-ligand complexes are available, it is still difficult to rationally engineer an
allosterically regulatable enzyme without decreasing its catalytic activity. Here, we describe an effective strategy to
deregulate the allosteric inhibition of enzymes based on the molecular evolution and physicochemical characteristics of
allostericligand-bindingsites.Wefoundthatallostericsitesareevolutionarilyvariableandcomprisedofmorehydrophobic
residuesthancatalyticsites.Weappliedourfindingstodesignmutationsinselectedtargetresiduesthatderegulatethe
allostericactivityoffructose-1,6-bisphosphatase(FBPase).Specifically,chargedaminoacidsatlessconservedpositionswere
substitutedwithhydrophobicorneutralaminoacidswithsimilarsizes.Theengineeredproteinssuccessfullydiminishedthe
allostericinhibitionofE.coliFBPasewithoutaffectingitscatalyticefficiency.Weexpectthatourmethodwillaidtherational
designofenzymeallostericregulationstrategiesandfacilitatethecontrolofmetabolicflux.
Citation: Yang J-S, Seo SW, Jang S, Jung GY, Kim S (2012) Rational Engineering of EnzymeAllosteric Regulation through Sequence Evolution Analysis. PLoS
ComputBiol8(7):e1002612.doi:10.1371/journal.pcbi.1002612
Editor:SarahA.Teichmann,MRCLaboratoryofMolecularBiology,Uni
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