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Amyloidogenesis of SARS-CoV-2 Spike Protein

• December 2021

DOI:10.1101/2021.12.16.472920

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Authors:

We Sofie Nystrom

• Linköping University

Per Hammarström

• Linköping University

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References (26)

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Abstract and Figures

SARS-CoV-2 infection is associated with a surprising number of morbidities. Uncanny similarities with amyloid-disease associated blood coagulation and fibrinolytic disturbances together with neurologic and cardiac problems led us to investigate the amyloidogenicity of the SARS-CoV-2 Spike protein (S-protein). Amyloid fibril assays of peptide library mixtures and theoretical predictions identified seven amyloidogenic sequences within the S-protein. All seven peptides in isolation formed aggregates during incubation at 37°C. Three 20-amino acid long synthetic Spike peptides (sequence 191-210, 599-618, 1165-1184) fulfilled three amyloid fibril criteria: nucleation dependent polymerization kinetics by ThT, Congo red positivity and ultrastructural fibrillar morphology. Full-length folded S-protein did not form amyloid fibrils, but amyloid-like fibrils with evident branching were formed during 24 hours of S-protein co-incubation with the protease neutrophil elastase (NE) in vitro. NE efficiently cleaved S-protein rendering exposure of amyloidogenic segments and accumulation of the peptide 193-202, part of the most amyloidogenic synthetic Spike peptide. NE is overexpressed at inflamed sites of viral infection and at vaccine injection sites. Our data propose a molecular mechanism for amyloidogenesis of SARS-CoV-2 S-protein in humans facilitated by endoproteolysis. The potential implications of S-protein amyloidogenesis in COVID-19 disease associated pathogenesis and consequences following S-protein based vaccines should be addressed in understanding the disease, long COVID-19, and vaccine side effects.

 

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1

Amyloidogenesis of SARS-CoV-2 Spike Protein

SofieNyström*‡,PerHammarström*‡

DeptofPhysics,ChemistryandBiology,LinköpingUniversity,Linköping,Sweden



ABSTRACT:SARS‐CoV‐2infectionisassociatedwithasurprisingnumberofmorbidities.Uncanny

similaritieswithamyloid‐diseaseassociatedbloodcoagulationandfibrinolyticdisturbancesto‐

getherwithneurologicandcardiacproblemsledustoinvestigatetheamyloidogenicityofthe

SARS‐CoV‐2Spikeprotein(S‐protein).Amyloidfibrilassaysofpeptidelibrarymixturesandtheoret‐

icalpredictionsidentifiedsevenamyloidogenicsequenceswithintheS‐protein.Allsevenpeptides

inisolationformedaggregatesduringincubationat37°C.Three20‐aminoacidlongsynthetic

Spikepeptides(sequence191‐210,599‐618,1165‐1184)fulfilledthreeamyloidfibrilcriteria:nu‐

cleationdependentpolymerizationkineticsbyThT,Congoredpositivityandultrastructuralfibrillar

morphology.Full‐lengthfoldedS‐proteindidnotformamyloidfibrils,butamyloid‐likefibrilswith

evidentbranchingwereformedduring24hoursofS‐proteinco‐incubationwiththeproteaseneu‐

trophilelastase(NE)invitro.NEefficientlycleavedS‐proteinrenderingexposureofamyloidogenic

segmentsandaccumulationofthepeptide193‐202,partofthemostamyloidogenicsynthetic

Spikepeptide.NEisoverexpressedatinflamedsitesofviralinfectionandatvaccineinjectionsites.

OurdataproposeamolecularmechanismforamyloidogenesisofSARS‐CoV‐2S‐proteininhumans

facilitatedbyendoproteolysis.ThepotentialimplicationsofS‐proteinamyloidogenesisinCOVID‐

19diseaseassociatedpathogenesisandconsequencesfollowingS‐proteinbasedvaccinesshould

beaddressedinunderstandingthedisease,longCOVID‐19,andvaccinesideeffects.

TheSARS‐CoV‐2pandemichasseverelyimpactedtheworldandofficialnumbersstatethatover270

millionpeoplehavebeeninfected(Dec2021),butunrecordedcasesarelikelysignificantlyhigher.Co‐

ronavirusesusethesurfacespikeprotein(S‐protein)toattachtohumancells.TheS‐proteinisaho‐

motrimerandeachsubunitofSARS‐CoV‐2S‐proteincomprise1273aminoacids.Fourcommoncorona

viruses;OC43,229E,NL63,HKU1infecthumansandcolonizetherespiratorytract.Recentlyemerged

SARS,MERSandsince2019alsoSARS‐CoV‐2,renderseveredisease.Althoughcoronavirusinfections

arecommon,ithasnotbeforeCOVID‐19beenreportsofsuchawidedistributionofcomplexsymptoms

involvingotherorgansthantherespiratorytract.Bloodclotting,heartfailure,peripheralneuropathy

andCNSdisordersareseveresymptomsoutofmanyreported.Whatcouldbethebasisforthispatho‐

genesis?Amyloidosismanifestsassystemicandlocalizeddisorderswithmanyoverlappingphenotypes

withreportedCOVID‐19symptoms.Wethereforehypothesizedonapotentialmolecularlink.Wewere

inspiredbyprevioushypothesesabouthumanandviralproteinamyloidsandinteractionsbetween

them[1‐3],inparticularSARS‐CoVspikeproteins[4‐6].WeaskedthequestionifSARS‐CoV‐2S‐protein

isamyloidogenic?

MotivatedbyZhangetal2018[5]wefirstobtainedapoollibraryofspikepeptides,intendedfor

antibodyscreening.Thelibrarycomprised316peptides(deliveredintwosubpoolsof158peptides

each)derivedfromapeptidescan(15‐merswith11AAoverlap,SupportingInf.2)throughtheentire

SARS‐CoV‐2S‐protein(ProteinID:P0DTC2).Thelibrarywasassayedforinvitroamyloidfibrilformation.

Fibrilswereformedinbothpeptidesubpools(SupportingInf.1andSFig.1).Encouragedbytheresults

wemovedontogenerate20AApeptidesequencesfromthefull‐lengthSARS‐CoV‐2S‐protein.We

aimedtoaddressthemostamyloidogenicsequencesandusedtheWALTZ

(https://waltz.switchlab.org/)predictionalgorithm[7]toidentifysuchsegments(Table1).

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preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in

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Table 1. Amino acid sequences and properties of synthetic SARS-CoV-2 S-protein peptides

PeptideAminoacidsequence*MW

(Da)**pIThTkinCongo

RedUltrastructure

Spike191 FVFKNIDGYFKIYSKHTPIN24319.4++fibril

Spike259WTAGAAAYYVGYLQPRTFLLK23899.5‐ +fibril

Spike362 KKKGGGYSVLYNSASFSTFK216910.0‐ +amorphous

Spike532NLVKNKCVNFNFNGLTGTGV21399.3++amorphous

Spike599GTNTSNQVAVLYQDVNCTEV21553.7++fibril

Spike689 KKKRSVASQSIIAYTMSLGA213910.5‐ ‐ribbons

Spike1165LGDISGINASVVNIQKEIDR21414.6++fibril

*ResiduesassignedincolorindicatetheamyloidogenicsegmentsaspredictedbyWALTZ.Highlightedingray

arenon‐nativeaminoacidsintroducedforsolubility.**Theoreticalmass(Dalton).Massspectraofpeptides

inSFig.2.



ThesevenamyloidogenicsequencesweredistributedovertheentireS‐proteinandarenamedaccord‐

ingtothestartingpositionoftheS‐protein(Fig.1).Allbutone(Spike362)ofthepredictedsequences

areinbeta‐sheetconformationinthecryo‐EMstructureofSARS‐CoV‐2Sinitsclosedstate[8].



Figure1.A.ThestructureofoneprotomerofthetrimericSARS‐CoV‐2S‐proteininitsclosedstate,PDBcode:6VXX[8]

withthepredictedfullsequenceoftheamyloidogenicpeptideshighlightedinthesamecolorsasthepredictionsinTable

1.B.ConformationofpeptideswithinthefoldedS‐proteinincomparisonwithAlphaFold2modelsofthesyntheticpep‐

tides(Table1)

.



ThemembranespanningC‐terminalpartoftheprotein(Spike1165)isnotincludedinthestructure.

ResultsfromAlphafold2[9]ontheselectedpeptidesindicatethatthreepeptides;Spike259,Spike362

andSpike689,thatareinβ‐strandconformationinthefull‐lengthproteinareinhelicalconformation

asfreepeptides(Fig.1B).Oneofthepeptides(Spike599)waspredictedbyAlphafold2toberandom

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preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in

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3

coilandthemembranespanningpartwaspredictedtobeanα‐helix.Notethatthesequencesof

Spike362andSpike689havebeenN‐terminallymodifiedwithsolubilizingsequencestoenableproduc‐

tion.Alphafold2wasperformedonthesequencesinTabl e 1(Fig.1B).

Lyophilizedpeptidesweresolubilizedinhexa‐fluoroisopropanol(HFIP)andweredilutedinPBS‐buffer

(pH7.5)toafinalpeptideconcentrationof0.1mg/ml(10%HFIP)andmonitoredforinvitroamyloid

fibrilformationkineticsusingThT,Congoredbirefringence(CR)andnegativestaintransmissionelec‐

tronmicroscopy(TEM).Fibrilswereformedwithinafewhoursfrommostofthesyntheticpeptidesby

atleastonebutnotallassays(Table1,Fig.2).Spike191,Spike532andSpike1165fulfilledallouramyloid

criteria:sigmoidalThTkinetics,Congophilicityandfibrillarultrastructure(Fig.2,Tabl e 1).Inparticular,

Spike191formedexceptionallywell‐orderedfibrils(Fig.2C,Fig.3C).

Thesyntheticpeptideswerealsopreparedasamixtureofallsevenpeptideswithafinalconcentra‐

tionof0.1mg/mloftotalpeptide(concentrationofeachpeptide~0.014mg/ml).Themixtureofpep‐

tidesresultedinamyloidfibrilformationwithsigmoidalThTkineticssuggestinganucleationdependent

mechanism(Fig.3A).Thefibrilmorphologyfromthemixture(Fig.3B)mostcloselyresembledthatof

fibrilscomposedofthewell‐orderedSpike191(cf.Fig3B;Fig.3C),suggestingthatthispeptideisdom‐

inatingthefibrilstructuresinthemixture

WhatwouldbeaplausiblemechanismforS‐proteinfibrilformationinaSARS‐CoV‐2infectedpatient?

SARS‐CoV‐2S‐proteinisfairlystable(Tm>50°C)[10](seebelow)andwouldnotreadilydenaturespon‐

taneously.Also,suchalargeproteinwithcomplexfoldwillnoteasilymisfoldintoanamyloidstate.

However,proteolysisisanobviouscandidatemechanism.

Endoproteolysisofprecursorproteinsisawell‐knownmolecularinitiationmechanisminseveralam‐

yloidosesnotablyAlzheimer´sdisease(AβPP),BritishandDanishdementia(ABri/ADan),andFinnish

familialamyloidosis(AGel).Proteolysisofthefull‐lengthproteinisalsoevidentinmanyotheramyloid

diseasedepositsfromATTR,ALys,AA,ASem1[11,12].SARS‐CoV‐2S‐proteinisproteolyzedduringin‐

fectionbyhostfurin‐likeenzymesandbyserineproteasessuchasthetransmembraneprotease,serine

2(TMPRSS2),atthecellsurface[13].S‐proteinisfurtherproteolyzedduringinflammation.Neutrophils

arethedominatingclassofleucocytesandareoneofthefirstrespondersduringacuteinflammation.

Neutrophilsarerecruitedtothebronchoalveolarspaceofpatientsinfectedwitharangeofdifferent

respiratoryviruses,includingSARS‐CoV‐2[14].Neutrophilsactbothbyphagocytosisofopsonizedpath‐

ogensandbyextracellularreleaseofenzymessuchasneutrophilelastase(NE).NEisaserineprotease

coupledtoobstructivelungdiseasessuchascysticfibrosis,chronicobstructivepulmonarydisease

(COPD)[15]andalpha‐1‐antitrypsindeficiency[16].TheaminoacidsequenceofSARS‐CoV‐2S‐protein

wastestedwithinsilicoproteolyticcleavagebyNEusingExpasyPeptidecutter.Oneoftheresulting

peptides,Spike193‐212,wascloselymatchingpeptideSpike191,onlyframeshiftedby2aminoacids

(Supportinginformation3)implyingatestablehypothesis.

Wethereforesubjectedfull‐lengthSARS‐CoV‐2S‐proteintoNEcleavageinvitro.Wefirstdetermined

thattheS‐proteinwasfoldedbythermalunfoldingexperimentsbydifferentialscanningfluorimetry

(DSF)(Fig.4,SFig.3).





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preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in

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

Figure2.AmyloidfibrilassaysofSARS‐CoV‐2Speptides(0.1mg/ml).A.FibrilformationkineticsmonitoredbyThTfluo‐

rescence.B.Congoredstainingmicroscopywithopenandcrossedpolarizers.C.UltrastructurebynegativestainTEM

.



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preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in

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

Figure3.Amyloidfibrillationof7mixedSARS‐CoV‐2S‐peptides(totalconcentration0.1mg/ml).A.Fibrilformationkinet‐

icsmonitoredbyThTfluorescence.B.FibrillarstructuresbynegativestainTEM.C.Fibrillarstructuresbynegativestain

TEMofSpike191resemblingthemix

.





Figure4.S‐proteinproteolysisbyNErendersamyloid‐likefibrils.A.ThermostabilityoftheSARS‐CoV‐2S‐protein,B.NE

andC.S‐protein+NE,measuredbyDSF.ThedashedlineinCisthemathematicalsumofS‐proteinandNErespectively

fromAandBwhichdoesnotfittheexperimentaldata,supportingcleavageofS‐proteinbyNE.MALDI‐ToFspectraofC18

isolatedpeptidesofD.S‐protein,E.NE,andF.S‐protein+NE(6h,37°C).TEMmicrographsofG.S‐proteinalonedepicting

theexpectedtrimers[17]H.NEalone,andI.S‐proteinandNEco‐incubatedatpH8.4for24h,37°C.Clustersofamyloid‐

likebranchedfibrilswereformedintheco‐incubationexperimentinI.





S‐proteinshowedacomplexunfoldingtrajectorywithmultipletransitionsaround45‐65°C,andamajor

unfoldingtransitionwithamidpointofthermaldenaturation(Tm)79°C(Fig.4A,SFig3A),similarto

thatreportedforanotherfull‐lengthS‐proteinconstruct[17].S‐proteinrefoldeduponloweringthe

temperaturealbeitwithanon‐cooperativerefoldingtransition(Fig.4A).NEunfoldedirreversiblywith

atTmof59°C(Fig.4B).TheincubatedS‐protein+NEonlyshowedanobvioustransitionforNEanddid

notrefolduponloweringthetemperaturesuggestingthatS‐proteinhadbeencleavedbyNE(Fig.4C).

WeverifieddigestionbymassspectrometrywhereonlytheS‐protein+NEexperimentrevealedpep‐

tidepeaks(Fig.4D‐F).Mostimportantlywediscoveredamyloid‐likefibrilformationdependentonthis

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6

proteolyticcleavagebyTEM.NeitherNEnorSARS‐CoV‐2S‐proteinincubatedaloneformedfibrils(Fig.

4G‐H).Fibrilswereonlyfoundafterco‐incubationofthetwoproteins(Fig.4I).Thefibrilshadanunu‐

sualmorphologywithevidentbranching(Fig.4I).Thismorphologysuggestsinvolvementofproteolyt‐

icallynickedS‐proteinwithinthefibrilrenderingnodesforbranchingofdifferentamyloidogenicse‐

quences(Fig.4I,SFig.4).To understandhowNEcleavedS‐proteinweperformedLC‐MS/MSanalysisof

peptidesformedafter1minand6hofdigestionat2:1excessofNEoverS‐protein.Wemapped98

identifiedNEcleavagepeptides(STable1)ontheS‐proteinstructureandclassedtheseintothree

groups:i)formedafter1min(Fig.5A,B),ii)formedafter1minandstillpresentafter6h(Fig.5A,C),

andiii)onlypresentafter6h(Fig.5A,D).Initialcleavageandfurtherdigestion(groupi)occurredmainly

withintheS2domainwithabundantcleavageoftheHRdomainsandintheC‐terminalpartofS1.NTD

andRBDweremuchlessaffected(Fig.5A).Threepersistentpeptidesformedafterinitialcleavage

(groupii)originatedfromNTDandRBD.Severalpeptidesonlyformedafter6hofincubation(group

iii).Strikinglythepeptidefromthesegment193‐202(FKNIDGYFKI,includedinSpike191)waspartof

thisgroupandwashighlyabundantafter6h(STable2).Comparedtotheamyloidogenicsequences

threepeptidescontainingsegmentsfromthesewereformedasfreepeptides(Spike191,Spike259,

Spike1165)stillpresentafter6hoursofco‐incubation,twoweredigestedearlyanddisappeared

(Spike532,Spike689),andtwowerelikelystillresidentintheparentnickedS‐protein(Spike362,

Spike599).Hencetheobservedformedbranchedfibrilsislikelycomposedofamixoffibrilsinitiated

byanamyloidogenicpeptideseedrecruitingnickedS‐proteinforelongationandbranching.





Figure5.NEcleavagesiteswithinfull‐lengthfoldedS‐protein.A.OverallmapofpeptidesidentifiedbyLC‐MS/MSincor‐

relationtotheamyloidogenicSpikepeptides(c.f.Fig.1andTable1)andtheS‐proteindomainstructure.Arrowsindicate

thelimitsofthecryo‐EMstructure27‐1146.Afteri)1min(red),ii)1minandstillpersistentat6h(magenta),iii)only

presentafter6hofincubation(blue).B‐D.thesamecolorcodeforcleavedpeptidegroupi‐iiimappedontotheprotomer

Cryo‐EMS‐proteinstructurePDBcode6VXX

.





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7

Whataretheimplicationsofourfindings?COVID‐19pathogenesisismultifactorialandcomplex[18].

SevereCOVID‐19includeacuterespiratorydistresssyndrome(ARDS)fromsevereinnateimmunesys‐

teminflammatoryreactionsresultinginlungdamage[19];Cytokinestorm[20];Heartdamage,includ‐

inginflammationoftheheartmuscle;Kidneydamage;Neurologicaldamage;Damagetothecirculatory

systemresultinginpoorbloodflow;Long‐COVIDsymptomsincludepersistentemotionalillnessand

othermentalhealthconditionsresemblingneurodegenerativediseases[18].

IthasbeenproposedthatsevereinflammatorydiseaseincludingARDSincombinationwithSARS‐

CoV‐2proteinaggregationmightinducesystemicAAamyloidosis[21].Neurotropiccolonizationand

cross‐seedingofS‐proteinamyloidfibrilstoinduceaggregationofhumanendogenousproteinshas

beendiscussedinthecontextofneurodegeneration[4].Notably,bloodclottingassociatedwithextra‐

cellularamyloidoticfibrillaraggregatesinthebloodstreamhavebeenreportedinaffectedCOVID‐19

patients[22].HypercoagulationandimpairedfibrinolysisweredemonstratedinexperimentallyS‐pro‐

teinspikedbloodplasmafromhealthydonors[22].Similarly,amyloidosisisassociatedwithcerebral

amyloidangiopathy,bloodcoagulationdisruption,fibrinolyticdisturbance[23,24]andFXIIKal‐

likrein/Kininactivationandthromboinflammation[25],suggestingpotentiallinksbetweenamyloido‐

genesisofS‐proteinandCOVID‐19phenotypes.

Inconclusion,wehereinproposedarathersimplemolecularmechanismforhowSARS‐CoV‐2S‐pro‐

teinendoproteolysisbyNEcanrenderexceptionallyamyloidogenicS‐peptidessuchassegment193‐

202,andexposureofmultipleamyloidogenicsegmentsinproteolyticallynickedS‐protein.Itispossible

thatotheramyloidogenicpeptidesandS‐proteinnickedbyotherproteasescouldbeinvolved.We

foundthatallcommoncoronavirusesinfectinghumanscontainamyloidogenicsequences(SFig.5A).

Nonetheless,themagnitudeofthediverseCOVID‐19symptomswasnotpreviouslyreported.Theseg‐

ment193‐212isuniqueforandSARS‐CoV‐2(SFig.5B),whichincombinationwithacuteinflammation

couldexplaintheputativeCOVID‐19associatedamyloidosis.

Furthermore,overthecourseoftheyear2021over8billiondosesofCOVID‐19vaccineshavebeen

administeredworldwide.MostdoseshaveprovidedSARS‐CoV‐2S‐proteinasmainantigen.Arecent

casereportdescribestheserendipitousdiscoveryofamyloidformationinahumanpatientlocallyat

thesiteofvaccinationandinaproximallymphnodewithin24hoursofthefirstdoseofmRNAvaccine

codingforS‐protein[26].Neutrophilrecruitmentandactivationatthesiteofvaccinationisexpected.

ThelocalizedamyloidwasdetectedbytheAβamyloidPETtracer18F‐Florbetabenalsoknowntobind

toAL,AAandATTRcardiacamyloidfibrils.Thereactantallegedamyloidogenicproteininthevaccinated

patientwasnotidentified.WetestedthefluorescentamyloidligandsCN‐PiB(fluorescentbenzothia‐

zoleanalogueofPittsburghcompoundB)andDF‐9(fluorescentstilbeneanalogueofFlorbetaben)and

foundstrongfluorescenceresponsetowardsSpike191fibrilsinvitro(SFig.6),supportingthepossibility

thatvaccineinducedS‐proteinderivedamyloiddepositionwasdetectedinthehumanPETimaging

casestudy[26].

IfthehereinproposedmechanismforS‐proteinamyloidformationisassociatedwithreportedcar‐

diac,‐blood,‐andnervoussystemdisordersincertainvaccinatedindividuals[27]isnotknown,neither

arethelong‐termconsequences,butisstronglyrecommendedtobeinvestigatedinthiscontext.

AUTHOR INFORMATION

*CorrespondingAuthors

SofieNyström[email protected],PerHammarström,[email protected]

AuthorContributions

‡Theresearchwasperformed,andmanuscriptwaswrittenwithequalcontributionsofbothauthors.

Bothauthorshavegivenapprovaltothefinalversionofthemanuscript.

FundingSources

ThisresearchwasfinancedbyTheSwedishResearchCouncilgrant#2019‐04405).

.CC-BY-NC-ND 4.0 International licenseperpetuity. It is made available under a

preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in

The copyright holder for thisthis version posted December 17, 2021. ; https://doi.org/10.1101/2021.12.16.472920doi: bioRxiv preprint

8

ACKNOWLEDGMENT

WethankBjörnWallnerforgeneratingAlphafold2foldingpredictionsofthesyntheticpeptides,

XiongyuWuforsynthesisofCN‐PiB,andJunZhangforsynthesisofDF‐9.WeacknowledgetheuseLiU

core‐facilitiesProLinC,MedicalFacultyMicroscopyandProteomicscoreespeciallyMariaTurkinafor

collectionofexperimentaldata.

ABBREVIATIONS

CNS,centralnervoussystem;SARS‐CoV‐2,SevereAcuteRespiratorySyndromeCoronavirus‐2,S‐pro‐

tein;Spikeprotein,ThT,ThioflavinT;CN‐PiB,Cyano‐PittsburghcompoundB;TEM,transmissionelec‐

tronmicroscopy;DSF,Differentialscanningfluorimetry

REFERENCES

1.Munch,J.,etal.,Semen‐derivedamyloidfibrilsdrasticallyenhanceHIVinfection.Cell,2007.131(6):p.

1059‐71.

2.Tayeb‐Fligelman,E.,etal.,InhibitionofamyloidformationoftheNucleoproteinofSARS‐CoV‐2.bioRxiv,

2021.

3.Michiels,E.,F.Rousseau,andJ.Schymkowitz,Mechanismsandtherapeuticpotentialofinteractions

betweenhumanamyloidsandviruses.CellMolLifeSci,2021.78(6):p.2485‐2501.

4.Tavassoly,O.,F.Safavi,andI.Tavassoly,SeedingBrainProteinAggregationbySARS‐CoV‐2asa

PossibleLong‐TermComplicationofCOVID‐19Infection.ACSChemNeurosci,2020.11(22):p.3704‐

3706.

5.Zhang,S.M.,etal.,Identificationandapplicationofself‐bindingzipper‐likesequencesinSARS‐CoV

spikeprotein.IntJBiochemCellBiol,2018.101:p.103‐112.

6.Idrees,D.andV.Kumar,SARS‐CoV‐2spikeproteininteractionswithamyloidogenicproteins:Potential

cluestoneurodegeneration.BiochemBiophysResCommun,2021.554:p.94‐98.

7.Maurer‐Stroh,S.,etal.,Exploringthesequencedeterminantsofamyloidstructureusingposition‐

specificscoringmatrices.NatMethods,2010.7(3):p.237‐42.

8.Walls,A.C.,etal.,Structure,Function,andAntigenicityoftheSARS‐CoV‐2SpikeGlycoprotein.Cell,

2020.181(2):p.281‐292e6.

9.Jumper,J.,etal.,HighlyaccurateproteinstructurepredictionwithAlphaFold.Nature,2021.596(7873):

p.583‐589.

10.Upadhyay,V.,etal.,Receptorbinding,immuneescape,andproteinstabilitydirectthenaturalselection

ofSARS‐CoV‐2variants.JBiolChem,2021.297(4):p.101208.

11.Benson,M.D.,etal.,Amyloidnomenclature2020:updateandrecommendationsbytheInternational

SocietyofAmyloidosis(ISA)nomenclaturecommittee.Amyloid,2020.27(4):p.217‐222.

12.Sipe,J.D.,Amyloidproteins:thebetasheetconformationanddisease.2005,Weinheim:Wiley‐VCH;

[Chichester:JohnWiley,distributor].

13.Peacock,T.P.,etal.,ThefurincleavagesiteintheSARS‐CoV‐2spikeproteinisrequiredfortransmission

inferrets.NatMicrobiol,2021.6(7):p.899‐909.

14.Johansson,C.andF.C.M.Kirsebom,Neutrophilsinrespiratoryviralinfections.MucosalImmunol,2021.

14(4):p.815‐827.

15.Pandey,K.C.,S.De,andP.K.Mishra,RoleofProteasesinChronicObstructivePulmonaryDisease.Front

Pharmacol,2017.8:p.512.

16.Strnad,P.,N.G.McElvaney,andD.A.Lomas,Alpha1‐AntitrypsinDeficiency.NEnglJMed,2020.

382(15):p.1443‐1455.

17.Edwards,R.J.,etal.,ColdsensitivityoftheSARS‐CoV‐2spikeectodomain.NatStructMolBiol,2021.

28(2):p.128‐131.

18.Huang,C.,etal.,6‐monthconsequencesofCOVID‐19inpatientsdischargedfromhospital:acohort

study.Lancet,2021.397(10270):p.220‐232.

19.Lipcsey,M.,etal.,TheOutcomeofCriticallyIllCOVID‐19PatientsIsLinkedtoThromboinflammation

DominatedbytheKallikrein/KininSystem.FrontImmunol,2021.12:p.627579.

20.Gao,Y.M.,etal.,Cytokinestormsyndromeincoronavirusdisease2019:Anarrativereview.JIntern

Med,2021.289(2):p.147‐161.

21.Sinha,N.andA.K.Thakur,LikelihoodofamyloidformationinCOVID‐19‐inducedARDS.Trends

Microbiol,2021.29(11):p.967‐969.

.CC-BY-NC-ND 4.0 International licenseperpetuity. It is made available under a

preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in

The copyright holder for thisthis version posted December 17, 2021. ; https://doi.org/10.1101/2021.12.16.472920doi: bioRxiv preprint

9

22.Grobbelaar,L.M.,etal.,SARS‐CoV‐2spikeproteinS1inducesfibrin(ogen)resistanttofibrinolysis:

implicationsformicroclotformationinCOVID‐19.BiosciRep,2021.41(8).

23.Hammarstrom,P.,Thebloodypathofamyloidsandprions.JThrombHaemost,2007.5(6):p.1136‐8.

24.Bouma,B.,etal.,Increasedplasmin‐alpha2‐antiplasminlevelsindicateactivationofthefibrinolytic

systeminsystemicamyloidoses.JThrombHaemost,2007.5(6):p.1139‐42.

25.Maas,C.,etal.,MisfoldedproteinsactivatefactorXIIinhumans,leadingtokallikreinformation

withoutinitiatingcoagulation.JClinInvest,2008.118(9):p.3208‐18.

26.Laudicella,R.,etal.,SubcutaneousUptakeon[18F]FlorbetabenPET/CT:aCaseReportofPossible

Amyloid‐BetaImmune‐ReactivityAfterCOVID‐19Vaccination.SNComprClinMed,2021:p.1‐3.

27.COVID‐19mRNAPfizer‐BioNTechVaccineAnalysisPrint.202117November2021[cited202129

November2021];Availablefrom:

https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/

1036445/COVID‐19_mRNA_Pfizer‐BioNTech_Vaccine_Analysis_Print_DLP_17.11.2021.pdf.



.CC-BY-NC-ND 4.0 International licenseperpetuity. It is made available under a

preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in

The copyright holder for thisthis version posted December 17, 2021. ; https://doi.org/10.1101/2021.12.16.472920doi: bioRxiv preprint

Citations (2)

References (26)

 

… With regard to viruses, in particular SARS-CoV2, the ability of its proteins and protein fragments to amyloidogenesis is unlikely to be an artifact. Interestingly, for the S-protein, the ability of its fragment to form amyloid-like fibrils was confirmed experimentally [27]. Also, the . …

Peptide from NSP7 is able to form amyloid-like fibrils: artifact or challenge to drug design?

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Effect of an Amyloidogenic SARS-COV-2 Protein Fragment on α-Synuclein Monomers and Fibrils

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• May 2022

• Asis K. Jana
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