Probing a Polar Cluster in the Retinal Binding Pocket of Bacteriorhodopsin by a Chemical Design Approach 英文参考文献.docVIP

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Probing a Polar Cluster in the Retinal Binding Pocket of Bacteriorhodopsin by a Chemical Design Approach 英文参考文献.doc

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Probing a Polar Cluster in the Retinal Binding Pocket of Bacteriorhodopsin by a Chemical Design Approach 英文参考文献

ProbingaPolarClusterintheRetinalBindingPocketof BacteriorhodopsinbyaChemicalDesignApproach RosanaSimo′n-Va′zquez1¤a,MartaDom?′nguez2,V?′ctorA.Lo′renz-Fonfr?′a1¤b,SusanaAlvarez′ 2,Jose′- Lu?′sBourdelande3,Angel′ R.deLera2,EstevePadro′s1,AlexPera′lvarez-Mar?′n1* 1DepartamentdeBioqu?′micaideBiologiaMolecularandCentred’EstudisenBiof?′sica,UniversitatAuto`nomadeBarcelona,Barcelona,Spain,2DepartamentodeQu?′mica Orga′nica.FacultaddeQu?′mica,UniversidadedeVigo,Vigo,Spain,3DepartamentodeQu?′mica(Qu?′micaOrga′nica)UniversitatAuto`nomadeBarcelona,Barcelona,Spain Abstract Bacteriorhodopsin has a polar cluster of amino acids surrounding the retinal molecule, which is responsible for light harvestingtofuelprotonpumping.Fromourpreviousstudies,wehaveshownthatthreonine90isthepivotalaminoacidin this polar cluster, both functionally and structurally. In an attempt to perform a phenotype rescue, we have chemically designedaretinalanaloguemoleculetocompensatethedrasticeffectsoftheT90Amutationinbacteriorhodopsin.This analogue substitutes the methyl group at position C13 of the retinal hydrocarbon chain by and ethyl group (20-methyl retinal).Wehaveanalyzedtheeffectofreconstitutingthewild-typeandtheT90Amutantapoproteinswithall-trans-retinal and its 20-methyl derivative (hereafter, 13-ethyl retinal). Biophysical characterization indicates that recovering the steric interactionbetweentheresidue90andretinal,easestheaccommodationofthechromophore,howeveritisnotenoughfor acompletephenotyperescue.Thecharacterizationofthesechemicallyengineeredchromoproteinsprovidesfurtherinsight into the role of the hydrogen bond network and the steric interactions involving the retinal binding pocket in bacteriorhodopsinandothermicrobialsensoryrhodopsins. Citation:Simo′n-Va′zquez R,Dom?′nguezM,Lo′renz-Fonfr?′aVA,A′lvarezS,BourdelandeJ-L,etal.(2012)ProbingaPolarClusterintheRetinalBindingPocketof BacteriorhodopsinbyaChemicalDesignApproach.PLoSONE7(8):e42447.doi:10.1371/journal.pone.0042447 Edito