Mitocryptides from Human Mitochondrial DNA–Encoded Proteins Activate Neutrophil Formyl Peptide Receptors Receptor Preference and Signaling Properties Michael Gabl.pdf

该【Mitocryptides from Human Mitochondrial DNA–Encoded Proteins Activate Neutrophil Formyl Peptide Receptors Receptor Preference and Signaling Properties Michael Gabl 】是由【小舍儿】上传分享，文档一共【15】页，该文档可以免费在线阅读，需要了解更多关于【Mitocryptides from Human Mitochondrial DNA–Encoded Proteins Activate Neutrophil Formyl Peptide Receptors Receptor Preference and Signaling Properties Michael Gabl 】的内容，可以使用淘豆网的站内搜索功能，选择自己适合的文档，以下文字是截取该文章内的部分文字，如需要获得完整电子版，请下载此文档到您的设备，方便您编辑和打印。:..MitocryptidesfromHumanMitochondrialDNA-EncodedProteinsActivateNeutrophilFormylPeptideReceptors:ReceptorPreferenceandSignalingPropertiesThisinformationiscurrentasofMarch30,,MartinaSundqvist,AndreHoldfeldt,SimonLind,JonasM?rtensson,KarinChristenson,TakayukiMarutani,ClaesDahlgren,HidehitoMukaiandHuameiForsmanJImmunolpublishedonline30March2018Downloadedfromtent/early/2018/03/29/?Submitonline.?RapidReviews!30days*fromsubmissiontoinitialdecision?NoTriage!Everysubmissionreviewedbypracticingscientists?FastPublication!eptancetopublicationbyguestonMarch30,2018*averageSubscriptionInformationaboutsubscribingtoTheJournalofImmunologyisonlineat:/subscriptionPermissionsSubmitcopyrightpermissionrequestsat:ut/Publications/JI/-:/alertsTheJournalofImmunologyispublishedtwiceeachmonthbyTheAmericanAssociationofImmunologists,Inc.,1451RockvillePike,Suite650,Rockville,MD20852Copyright?2018byTheAmericanAssociationofImmunologists,:0022-1767OnlineISSN:1550-6606.:..PublishedMarch30,2018,doi:?EncodedProteinsActivateNeutrophilFormylPeptideReceptors:ReceptorPreferenceandSignalingPropertiesMichaelGabl,*MartinaSundqvist,*AndreHoldfeldt,*SimonLind,*JonasMartensson,*?KarinChristenson,?TakayukiMarutani,?ClaesDahlgren,*HidehitoMukai,?andHuameiForsman*eutrophilsexpressformylpeptidereceptors(FPRs;FPR1andFPR2)thatdistinctlyrecognizepeptidesstartingwithanN-formylatedmethionine(fMet).Thisisahallmarkofbacterialmetabolism;similartoprokaryotes,thestartingaminoacidinsynthesisofmitochondrialDNA?(mitocryptides;MCTs)withanN-terminalfMetcouldbeidenti?edbyourinnateimmunesystem;however,incontrasttoourknowledgeaboutbacterialDownloadedfrommetabolites,verylittleisknownabouttherecognitionpro?,wedeterminedtheneutrophil-recognitionpro?lesandfunctionaloutputofputativeMCTsoriginatingfromtheNterminiofthe13humanmitochondrialDNA?-recognitionpro?les:MCTsfromND3andND6haveareceptorpreferenceforFPR1;MCTsfromtheproteinsND4,ND5,andcytochromebpreferFPR2;andMCT-COX1isadualFPR1/,whereasMCTsfromND1,ATP6,ATP8,COX2,andCOX3,,theactivatingMCTsheterologously/desensitizedIL-8Rbutprimedtheresponsetotheplatelet-,ourdatasuggestthatMCTshavebiasedsignalingpropertiesinfavorofactivationofthesuperoxide-generatingNADPHoxidaseorrecruitmentofb-,weidentifyseveralnovelFPR-activatingpeptideswithsequencespresentintheNterminiofmitochondrialDNA?encodedproteins,,2018,200:000?(neutrophilsandmonocytes)constitutecells(1,2).Thephagocytesareequippedwithrecognitionmoleculesasecuritysysteminperipheralbloodandtissueswherethey(receptors)thataredesignedtoidentifyandrespondtomicrobe-Pactasanimportant?rstlineofcellulardefensethatcanbeassociatedmolecularpatternsanddanger-associatedmolecularbyguestonMarch30,(DAMPs)(3).Amongthedifferentpatternrecognitionrecep-systemiswellconserved,adoptingrecognitionofmolecularpatternstorsexpressedinphagocytes,formylpeptidereceptors(FPRs)partici-totrackdowninvasionbymicrobialpathogens,aswellasimminentpateintherecruitmentofin?ammatorycellstositesofinfectionand/ordangersignalsreleasedduringtraumaticdestructionofhosttissues/tissueinjury(4,5).umulationatthesiteofabacterialinfectionhasbeenattributedtotherecognitionofbacterial-derivedN-formylatedpeptidesasmicrobe-associated*DepartmentofRheumatologyandIn?ammationResearch,UniversityofGothenburg,?molecularpatternsbyFPRs(6,7).Protein/peptidesynthesisstarting41390Gothenburg,Sweden;DepartmentofOralMicrobiologyandImmunology,InstituteofOdontology,SahlgrenskaAcademy,UniversityofGothenburg,41390Gothenburg,?withanN-formylatedmethionine(fMet)residueisahallmarkofbac-Sweden;andLaboratoryofPeptideScience,GraduateSchoolofBio-Science,Nagahamateriametabolism(8,9),andtheformylgroupissubsequentlycleavedInstituteofBio-ScienceandTechnology,526-0829Nagahama,JapanORCIDs:0000-0002-0859-0792(.);0000-0001-6469-4345(.).byadeformylasetogeneratematureproteins,oritisleftuntouchedonsecretedpeptides,suchasthephenol-solublemodulins,peptidesReceivedforpublicationDecember11,,usaureus(10).Ac-,theKingGustafVcordingly,partialinhibitionoftheformylmethioninedeformylase80-YearFoundation,theSwedishgovernmentundertheAvtalomLa¨karutbildningduringbacterialgrowthresultsinareleaseofincreasingamountsofochForskningagreement,theClasGroschinskyMemorialFoundation,theIngabrittformylpeptides(11–13).andArneLundbergFoundation,theWilhelmandMartinaLundgrensScienti?cFoundation,,andbyaresearch?FPR1wasthe?rstreceptoridenti?edthatrecognizesbacterial-grantfromtheMinistryofEducation,Culture,Sports,ScienceandTechnologyofderivedformylpeptideswithhighaf?nity;however,fairlyrecently,Japan(25350971).FPR2wasalsoshowntorecognizesuchpeptideswithhighaf?,Departmentof(4,14).FPR1andFPR2share69%sequenceidentityandhaveRheumatologyandIn?ammationResearch,Go¨University,Guldhedsgatan10A,41346Gothenburg,-mailaddress:huamei.******@,butsomewhatoverlapping,ligand-recognitionpro?lesAbbreviationsusedinthisarticle:ATP6,ATPsynthasesubunitprotein6;CL,isolumi-(5).Ithasbeensuggestedthattheligand-bindingpocketinFPR1nol-ECL;COX1,cytochromecoxidasesubunitI;CysH,cyclosporineH;DAMP,danger-hasroomfor4or5aa,includingthefMetresidue,butthepreciseassociatedmolecularpattern;fMet,formylatedmethionine;FPR,formylpeptidereceptor;2+herespectiverecognitionKRG,Krebs–Ringerphosphatebuffersup2+plementedwithglucose(10mM),Ca(1mM),andMg(,);MCT,mitochondrialcrypticpeptide(mitocryptide);pro?leshavenotbeende?ned(15).EarlierstudieshavedemonstratedND6,NADHdehydrogenasesubunit6;PAF,platelet-activatingfactor;PAFR,PAFthattheN-(4),butsequencesinmoredistantpartsoftheligandmayalsoCopyrightó2018byTheAmericanAssociationofImmunologists,-1767/18/$(16,17).Thisisde?nitelythecasefor/doi/:..2RECOGNITIONOFMITOCHONDRIAL-DERIVEDFORMYLPEPTIDESBYFPRsFPR2;itrecognizeslongerpeptidesinwhichresidueslocatedoutsideMCTsandpeptidesynthesisofthepresumedbindingpocketareofimportance(16,17).Theaminoacidsequencesofthe13humanmitochondrialDNA–encodedanizedasadouble-strandedproteinswereobtainedfromtheSwiss-Protdatabase(TableI).Theputativecircularmolecule,which,-formylatedpeptidesformedfromtheseproteins,followingproteolyticribosomalRNAandtRNAmolecules,encodesfor13proteinsthatarecleavagewithtrypsinorchymotrypsin,werepredictedusingthePeptide-ponentsoftherespiratorychaingeneratingthemajorityCuttertool(http://web./peptide_cutter/),withtheexceptionofNADHdehydrogenasesubunit6(ND6),becausethisisthoughttobelocatedintheofcellularATPviaoxidativephosphorylation(18).Beingofpro-,wetemporarilykaryoticorigin,mitochondrialDNAistranscribedandtranslatednamedthese13cleavagefragmentsindependentoftheiractivityasMCTsbyasystemindependentofthatusedfornuclearhostDNA,andforeasierunderstanding,although“mitocryptides”wereoriginallyde?nedastheinitiatingaminoacidinthemitochondrial-encodedproteinsneutrophil-activatingpeptidesthatarederivedfrommitochondrialproteins(30).ThecleavagesitesintheparentproteinstogeneratetheMCTsareasisanfMet,meaningthatthismicrobialhallmarkisalsovalidforfollows:ATPsynthasesubunitprotein6(ATP6;ataminoacidposition9),mitochondrial-encodedproteins(19,20).TissuedestructionandATP8(ataminoacidposition12),cytochromecoxidasesubunitI(COX1;atcellnecrosiswillultimatelyleadtoareleaseofotherwisehidden/aminoacidposition13),COX2(ataminoacidposition23),COX3(ataminocrypticendogenousdangermolecules,includingmitochondrialacidposition12),ND1(ataminoacidposition10),ND2(ataminoacidposition10),ND3(ataminoacidposition5),ND4(ataminoacidposition20),crypticpeptides(mitocryptides;MCTs)(21,22).InadditiontoND4L(ataminoacidposition5),ND5(ataminoacidposition28),andMCTs,mitochondriacontainATPandmitochondrialDNA,mol-ND6(ataminoacidposition6)(seeaminoacidsequenceinTableI).MCT-2eculesthatallpotentiallycanactasDAMPsandinduceaproin-derivedfromhumancytochromeb(ataminoacidposition15)waspre-?ammatoryresponse(19,23,24).Therefore,itisreasonabletodictedbyhomologytotheearlierdescribedporcineMCT-2(28).Allofthesepeptideswerechemicallysynthesizedbyasolid-phasemethodusingassumethatreceptorsrecognizingthefMet–peptidepatternhave9-?uorenylmethyloxycarbonylandpuri?edbyreverse-phasehigh-performanceDownloadedfromevolvedtoclearandresolveinfectiousandasepticin?ammatoryliquidchromatography,andtheirhomogeneitywascon?rmedbyreverse-,mitochondrial-derivedhigh-,hence,AllpeptidesweredissolvedinDMSOtoaconcentrationof10mMandstoredat280?-Ringer2+induceasepsis-likestateduringsevereinjury,althoughthechemicalphosphatebuffersupplementedwithglucose(10mM),Ca(1mM),andstructuresofthosemoleculeshavenotbeendetermined(25).Further,Mg2+(,)(KRG).mitochondrialDAMPshavealsobeendemonstratedtobeofim-/portancefortherecruitmentofinnateimmunecellstoasepticin-Chemicals?ammatorysites(26,27).Hence,anunderstanding,atthemolecularThehexapeptideWKYMVM(agonistforFPR2)wasfromAltaBiosciencelevel,ofinnateimmunerecognitionofmitochondrial-derivedDAMPs,(UniversityofBirmingham,Birmingham,.).TheformylpeptidefMLFincludingtheformylpeptides,an(theprototypeagonistforhumanFPR1),IL-8,DMSO,BSA,andisoluminolwerepurchasedfromSigma-Aldrich(,MO).Thereceptorantagonistfailureandsevereinjury/in?,MCT-2,forFPR1,cyclosporineH(CysH),waskindlyprovidedbyNovartisPharmaa15-residue-longformylpeptidewithasequenceidenticaltothe(Basel,Switzerland),andthereceptorantagonistforFPR2,PBP10[-terminusofmitochondrialDNA–encodedcytochromeb,isthe31fordetailsaboutthisinhibitorymolecule],wasobtainedfromCalbiochemonlyformylpeptidethathasbeenisolatedfromhosttissue(28,29).(SanDiego,CA).Allophycocyaninin-andPE-conjugatedAbsagainstCD62LandCD11bwerefromBectonDickinson(SanJose,CA),andFITC-conjugatedbyguestonMarch30,2018Thefactthat12otherproteins,inadditiontocytochromeb,areAbagainstCD66bwasfromAbDSerotec/Bio-Rad(Sundbyberg,Sweden).encodedforbymitochondrialDNAsuggeststhatotherendoge-rTNF-awasfromR&DSystemsEurope(Abingdon,Oxon,.),andHRPnousN-formylatedMCTsmaybeformedduringtissuedestruction/wasfromBoehringerMannheim(Mannheim,Germany).DextranandFicoll-necrosisandhaveabilitytointeractwiththeinnateimmunesystemPaquewereobtainedfromGELifeSciences(Uppsala,Sweden).(20,30).IsolationofhumanneutrophilsInanattempttogainmolecularinsightsintotheinnateimmune–modulationpropertiesofendogenousin?ammatorymediatorsofHumanperipheralbloodneutrophilswereisolatedfrombuffycoatsorfromfreshlydrawnbloodofhealthydonorsusingdextransedimentationandmitochondrialorigin,wesynthesized13putativeformylatedMCTsFicoll-Paquegradientcentrifugation,asdescribed(32).Theremainingeryth-withpeptidesequencesidenticaltotheNterminiofproteinsencodedrocytesweredisruptedbyhypotoniclysisandtheneutrophilswerewashed,forbyhumanmitochondrialDNA(TableI).Theireffectsonneu-resuspendedinKRG,?edwithminim