Protein Glycosylation in Helicobacter pylori Beyond the Flagellins 英文参考文献.docVIP

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Protein Glycosylation in Helicobacter pylori Beyond the Flagellins 英文参考文献.doc

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Protein Glycosylation in Helicobacter pylori Beyond the Flagellins 英文参考文献

ProteinGlycosylationinHelicobacterpylori:Beyondthe Flagellins? PatrickS.Hopf,RachelS.Ford,NajwaZebian,AlexandraMerkx-Jacques,SomalingaVijayakumar, DinathRatnayake,JacquelineHayworth,CaroleCreuzenet* InfectiousDiseasesResearchGroup,DepartmentofMicrobiologyandImmunology,TheUniversityofWesternOntario,London,Ontario,Canada Abstract GlycosylationofflagellinsbypseudaminicacidisrequiredforvirulenceinHelicobacterpylori.Wedemonstratethat,inH. pylori,glycosylationextendstoproteinsotherthanflagellinsandtosugarsotherthanpseudaminicacid.Severalcandidate glycoproteinsdistinctfromtheflagellinsweredetectedviaProQ-emeraldstainingandDIG-orbiotin-hydrazidelabelingof thesolubleandoutermembranefractionsofwild-typeH.pylori,suggestingthatproteinglycosylationisnotlimitedtothe flagellins. DIG-hydrazide labeling of proteins from pseudaminic acid biosynthesis pathway mutants showed that the glycosylation of some glycoproteins is not dependent on the pseudaminic acid glycosylation pathway, indicating the existenceofanovelglycosylationpathway.Fractionsenrichedinglycoproteincandidatesbyionexchangechromatography were used to extract the sugars by acid hydrolysis. High performance anion exchange chromatography with pulsed amperometricdetectionrevealedcharacteristicmonosaccharidepeaksintheseextracts.Themonosaccharideswerethen identifiedbyLC-ESI-MS/MS.Thespectraareconsistentwithsugarssuchas5,7-diacetamido-3,5,7,9-tetradeoxy-L-glycero-L- manno-nonulosonic acid (Pse5Ac7Ac) previously described on flagellins, 5-acetamidino-7-acetamido-3,5,7,9-tetradeoxy-L- glycero-L-manno-nonulosonic acid (Pse5Am7Ac), bacillosamine derivatives and a potential legionaminic acid derivative (Leg5AmNMe7Ac)whichwerenotpreviouslyidentifiedinH.pylori.Thesedataopenthewaytothestudyofthemechanism androleofproteinglycosylationonproteinfunctionandvirulenceinH.pylori. Citation: Hopf PS, Ford RS, Zebian N, Merkx-Jacques A, Vijayakumar S, et al. (2011) Protein Glycosylation in Helicobacter pylori: Beyond the Flagellins? PLoS ONE6(9):