Sequence Conservation in Plasmodium falciparum α-Helical Coiled Coil Domains Proposed for Vaccine Development 英文参考文献.docVIP

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Sequence Conservation in Plasmodium falciparum α-Helical Coiled Coil Domains Proposed for Vaccine Development 英文参考文献.doc

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SequenceConservationinPlasmodiumfalciparuma- HelicalCoiledCoilDomainsProposedforVaccine Development CarolineKulangara1,AndreyV.Kajava2,GiampietroCorradin3,IngridFelger1* 1SwissTropicalInstitute,Basel,Switzerland,2CentredeRecherchesdeBiochimieMacromoleculaire,FRE-2593CNRS,Montpellier,France,3DepartmentofBiochemistry, UniversityofLausanne,Epalinges,Switzerland Abstract Background:TheavailabilityoftheP.falciparumgenomehasledtonovelwaystoidentifypotentialvaccinecandidates.A new approach for antigen discovery based on the bioinformatic selection of heptad repeat motifs corresponding to a- helicalcoiledcoilstructuresyieldedpromisingresults.Toelucidatethequestionabouttherelationshipbetweenthecoiled coilmotifsandtheirsequenceconservation,wehaveassessedtheextentofpolymorphisminputativea-helicalcoiledcoil domainsinculturestrains,innaturalpopulationsandinthesinglenucleotidepolymorphismdataavailableatPlasmoDB. Methodology/Principal Findings: 14 a-helical coiled coil domains were selected based on preclinical experimental evaluation.TheyweretestedbyPCRamplificationandsequencingofdifferentP.falciparumculturestrainsandfieldisolates. We found that only 3 out of 14 a-helical coiled coils showed point mutations and/or length polymorphisms. Based on promisingimmunologicalresults5ofthesepeptideswereselectedforfurtheranalysis.Directsequencingoffieldsamples from Papua New Guinea and Tanzania showed that 3 out of these 5 peptides were completely conserved. An in silico analysis of polymorphism was performed for all 166 putative a-helical coiled coil domains originally identified in the P. falciparumgenome.Wefoundthat82%(137/166)ofthesepeptideswereconserved,andforonepeptideonlythedetected SNPs decreasedsubstantially the probability score for a-helicalcoiled coil formation.MoreSNPs were foundinarrays of almostperfecttandemrepeats.Insummary,thecoiledcoilstructurepredictionwasrarelymodifiedbySNPs.Theanalysis revealedanumberofpeptideswithstrictlyconserveda-helicalcoiledcoilmotifs. Conclusio