Functional Characterization of a Newly Identified Group B Streptococcus Pullulanase Eliciting Antibodies Able to Prevent Alpha-Glucans Degradation 英文参考文献.docVIP

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Functional Characterization of a Newly Identified Group B Streptococcus Pullulanase Eliciting Antibodies Able to Prevent Alpha-Glucans Degradation 英文参考文献.doc

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Functional Characterization of a Newly Identified Group B Streptococcus Pullulanase Eliciting Antibodies Able to Prevent Alpha-Glucans Degradation 英文参考文献

FunctionalCharacterizationofaNewlyIdentifiedGroup BStreptococcusPullulanaseElicitingAntibodiesAbleto PreventAlpha-GlucansDegradation IsabellaSanti.,AlfredoPezzicoli.,MattiaBosello,FrancescoBerti,MassimoMariani,JohnL.Telford, GuidoGrandi,MarcoSoriani* NovartisVaccinesandDiagnosticsSrl,Siena,Italy Abstract Streptococcal pullulanases have been recently proposed as key components of the metabolic machinery involved in bacterialadaptation tohost niches.By sequenceanalysis oftheGroup B Streptococcus (GBS)genomewefounda novel putativesurfaceexposedproteinwithpullulanaseactivity.WenamedsuchaproteinSAP.Thesapgeneishighlyconserved amongGBSstrainsandhomologousgenes,suchasPulAandSpuA,havebeendescribedinotherpathogenicstreptococci. The SAP protein contains two N-terminal carbohydrate-binding motifs, followed by a catalytic domain and a C-terminal LPXTGcellwall-anchoringdomain.InvitroanalysisrevealedthattherecombinantformofSAPisabletodegradea-glucan polysaccharides, such as pullulan, glycogen and starch. Moreover, NMR analysis showed that SAP acts as a type I pullulanase. Studies performed on whole bacteria indicated that the presence of a-glucan polysaccharides in culture medium up-regulated the expression of SAP on bacterial surface as confirmed by FACS analysis and confocal imaging. Deletionofthesapgeneresultedinareducedcapacityofbacteriatogrowinmediumcontainingpullulanorglycogen,but not glucose or maltose, confirming the pivotal role of SAP in GBS metabolism of a-glucans. As reported for other streptococcalpullulanases,wefoundspecificanti-SAPantibodiesinhumanserafromhealthyvolunteers.Investigationof thefunctionalroleofanti-SAPantibodiesrevealedthatincubationofGBSinthepresenceofserafromanimalsimmunized withSAPreducedthecapacityofthebacteriumtodegradepullulan.Ofinterest,anti-SAPsera,althoughtoalowerextent, alsoinhibitedGroupAStreptococcuspullulanaseactivity.ThesedataopennewperspectivesonthepossibilitytouseSAP asapotentialvaccinecomponentinducingfunctionalcross-reactinganti