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Amyloidogenesis of SARS-CoV‑2 Spike Protein
Soﬁe Nyström*,† and Per Hammarström*,†Cite This: J. Am. Chem. Soc. 2022, 144, 8945−8950

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sı Supporting Information*

ABSTRACT: SARS-CoV-2 infection is associated with a surprising number of morbidities. Uncanny similarities with amyloiddisease 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 theoreticalpredictions 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 192−211, 601−620, 1166−1185) fulﬁlled threeamyloid 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 h of S-protein coincubation with the protease neutrophil elastase (NE) in vitro. NE eﬃciently cleaved S-protein, rendering
exposure of amyloidogenic segments and accumulation of the amyloidogenic peptide 194−203, part of the most amyloidogenic
synthetic spike peptide. NE is overexpressed at inﬂamed sites of viral infection. Our data propose a molecular mechanism forpotential amyloidogenesis of SARS-CoV-2 S-protein in humans facilitated by endoproteolysis. The prospective of S-protein
amyloidogenesis in COVID-19 disease associated pathogenesis can be important in understanding the disease and long COVID-19.

C

Amyloidosis is associated with cerebral amyloid angiopathy,
blood coagulation disruption, ﬁbrinolytic disturbance,8,9 FXII
Kallikrein/Kinin activation, and thromboinﬂammation,10 suggesting potential links between S-protein amyloidogenesis and
COVID-19 phenotypes. We therefore hypothesized a potential
molecular link between S-protein and amyloid formation.
Inspired by previous hypotheses about human and viral protein
amyloids and interactions between them,11−13 in particular
SARS-CoV spike proteins,6,14,15 we asked the question: Is SARSCoV-2 S-protein amyloidogenic?
We obtained a 316 peptide pool library (divided into two
subpools) from a peptide scan through the entire SARS-CoV-2
S-protein (ProteinID: P0DTC2) (Supporting Information). In
vitro amyloid ﬁbrils were formed in both peptide subpools(Supporting Information, Figure S1). Encouraged by the results,
we generated 20-AA peptides 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 algorithm16 to identify such segments (Table 1,
Supporting Information).
Seven amyloidogenic sequences distributed over the entire Sprotein were identiﬁed and named according to the starting
position in the S-protein (Figure S2, Supporting Information).
All but one (Spike365) of the predicted sequences are in β-sheet
conformation in the SARS-CoV-2 Spike cryo-EM structure in its

oronaviruses use the homotrimeric surface spike protein(S-protein) to attach to human cells. Each SARS-CoV-2 Sprotein subunit comprises 1273 amino acids.1 Four common
coronaviruses (OC43, 229E, NL63, and HKU1) infect humans
and colonize the respiratory tract. Recently emerged SARS,MERS, and since 2019 also SARS-CoV-2 result in severe disease.
Although coronavirus infections are common, not before
COVID-19 has such a wide distribution of complex symptoms
involving organs other than the respiratory tract been reported.
COVID-19 pathogenesis is multifactorial and complex.2 Severe
COVID-19 includes acute respiratory distress syndrome(ARDS) from innate immune system inﬂammatory reactions
resulting in lung damage;3 cytokine storm;4 heart damage,including heart muscle inﬂammation; kidney damage; neurological damage; damage to the circulatory system resulting inpoor blood ﬂow. Long-COVID-19 symptoms include persistentemotional illness and mental health conditions resemblingneurodegenerative diseases.2 What could be the basis for thispathogenesis?
Amyloidosis from several culprit proteins manifests assystemic and localized disorders with many phenotypes
overlapping with reported COVID-19 symptoms. It has been
proposed that severe inﬂammatory disease including ARDS in
combination with SARS-CoV-2 protein aggregation might
induce systemic AA amyloidosis.5 Neurotropic colonizationand cross-seeding of S-protein amyloid ﬁbrils to induce
aggregation of endogenous proteins has been discussed in thecontext of neurodegeneration. 6 Notably, blood clotting
associated with extracellular amyloidotic ﬁbrillar aggregates inthe bloodstream has been reported in COVID-19 patients.7
Hypercoagulation/impaired ﬁbrinolysis was demonstrated inblood plasma from healthy donors experimentally spiked with Sprotein.7© 2022 The Authors. Published byAmerican Chemical Society

Received: April 13, 2022
Published: May 17, 2022

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Table 1. Amino Acid Sequences and Properties of Synthetic SARS-CoV-2 S-Protein peptidesaResidues assigned in color indicate the amyloidogenic segments as predicted by WALTZ. Highlighted in gray are non-native amino acids
introduced for solubility. bTheoretical mass.

Fibrils from most of the synthetic peptides were detected
within a few hours by at least one assay (Table 1, Figure 1).
Spike192, Spike601, and Spike1166 fulﬁlled all our amyloid
criteria: sigmoidal ThT kinetics, Congophilicity, and ﬁbrillar
ultrastructure (Figure 1, Table 1). Spike192 formed exceptionally well-ordered ﬁbrils comparable to a mix of all peptides(Figures 1C and S3C).
What would be a plausible mechanism for S-protein ﬁbril
formation during SARS-CoV-2 infection? SARS-CoV-2 Sprotein is fairly stable (Tm > 50 °C)17 and would not readily
denature spontaneously. Furthermore, such a large protein withcomplex folding 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 amyloidoses, notably
Alzheimer′s disease (AβPP), British and Danish dementia(ABri/ADan), and Finnish familial amyloidosis (AGel).
Proteolysis of full-length proteins is also evident in many otheramyloid disease deposits from ATTR, ALys, AA, andASem1.18,19
SARS-CoV-2 S-protein is proteolyzed during infection by
host furin-like enzymes and by serine proteases such as thetransmembrane protease, serine 2 (TMPRSS2), at the cellsurface20 and is further proteolyzed during inﬂammation.
Neutrophils are the dominating class of leukocytes and a ﬁrst
responder during acute inﬂammation. Neutrophils are recruited
to the bronchoalveolar space of patients infected with a range ofdiﬀerent respiratory viruses, including SARS-CoV-2.21 Neutrophils act by phagocytosis of opsonized pathogens and by
extracellular release of enzymes such as neutrophil elastase(NE). NE is a serine protease coupled to obstructive lungdiseases such as cystic ﬁbrosis, chronic obstructive pulmonary
disease,22 and alpha-1-antitrypsin deﬁciency.23
The amino acid sequence of SARS-CoV-2 S-protein was
subjected to in silico proteolytic cleavage by NE using Expasy
Peptide cutter. One of the resulting peptides, Spike194−213,
closely matched Spike192, only frame shifted by two aminoacids (Supporting Information), implying a testable hypothesis.
We subjected full-length SARS-CoV-2 S-protein to NE
cleavage in vitro. S-protein showed a complex thermal unfolding
trajectory with multiple transitions around 45−65 °C and amajor unfolding transition with a midpoint of denaturation (Tm)of 79 °C (Figures 2A and S4A) with diﬀerential scanning
ﬂuorimetry (DSF) (Figures 2 and S4), similar to literature values
for full-length S-proteins,24 conﬁrming folded protein at starting
point. S-protein refolded upon cooling albeit noncooperatively(Figure 2A). NE unfolded irreversibly (Tm 59 °C) (Figure 2B).
Co-incubated S-protein+NE only showed an obvious transition
for NE and did not refold upon cooling, suggesting that Sprotein had been cleaved by NE (Figure 2C). Mass

Figure 1. Amyloid ﬁbril assays of SARS-CoV-2 S peptides (0.1 mg/mL). (A) ThT ﬂuorescence ﬁbril formation kinetics. (B) Congo redbirefringence microscopy. (C) Negative stain TEM ultrastructure.

closed state.1 The C-terminal part of the protein (Spike1166) is
not resolved in the structure.
Solubilized peptides (0.1 mg/mL, PBS pH 7.5, 10% HFIP)
were monitored for in vitro amyloid ﬁbril formation kinetics
using ThT, Congo red birefringence (CR), and negative staintransmission electron microscopy (TEM).
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Figure 2. S-protein proteolysis by NE renders amyloid-like ﬁbrils. Thermostability of (A) SARS-CoV-2 S-protein, (B) NE, (C) S-protein+NE,
measured by DSF. Dashed line in (C) is the mathematical sum of S-protein and NE, respectively, from (A) and (B) 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, (H) NE alone, and (I) S-protein+NE coincubated at pH 8.4, 24 h, 37 °C.

S-protein (Spike365 and Spike601). Hence, the observed
formed branched ﬁbrils (Figures 2I and S5) are likely composedof a mix of ﬁbrils initiated by an amyloidogenic peptide seedrecruiting nicked S-protein for elongation and branching.
We performed ﬁbrillation experiments identical to that for theother spike peptides also on the short fragment 194−203(Figure 4A, B). The peptide was less amyloidogenic than
Spike192 (1 mg/mL was required to form ﬁbrils compared to
0.1 mg/mL for Spike192) (Figure 4B). Formed ﬁbrils of
Spike194−203 were however amyloidogenic by our threecriteria, ThT kinetics, Congo red birefringence, and ﬁbrillar
ultrastructure (Figure 4A, B). It is worth noting that the
Spike194−203 peptide lacks one amino acid in the predicted
amyloidogenic sequence. To test the signiﬁcance of this aminoacid deletion in the peptide, we performed a simple in silico
mutation experiment where we substituted the ﬁnal tyrosine in
the amyloidogenic segment of Spike192 with a glycine to mimic
its deletion. The in silico substitution Y204G abolished theamyloid prediction (Figure S6), demonstrating that removal ofthis amino acid will alter the amyloidogenic properties of thepeptide. Fibril “shaving” is known for several amyloidogenic
proteins and peptides.25 It is possible that Y204 is cleaved after
S-protein aggregation.
It is known that S-protein aﬀects the formation of persistentamyloid-like microclots in human blood, a potential pathologicalcause of long COVID-19 symptoms.26 We performed athrombin induced ﬁbrinogen to ﬁbrin conversion followed byplasminogen tPA assay27 in the presence and absence of Spike
peptide ﬁbrils (Supporting Information).
The addition of 10 μg/mL amyloid ﬁbrils formed from a mixof the seven spike peptides (Table 1, Figure S3) during ﬁbrin

spectrometry veriﬁed digestion since only the S-protein+NE
experiment revealed peptide peaks (Figure 2D−F).
Most importantly, we discovered amyloid-like ﬁbril formation
upon proteolytic cleavage using TEM. Neither NE nor SARSCoV-2 S-protein incubated alone formed ﬁbrils (Figure 2G−H).
Fibrils were found only after co-incubation of the two proteins(Figure 2I). The ﬁbrils showed unusual morphology withevident branching (Figure 2I), suggesting involvement ofproteolytically nicked S-protein within the ﬁbril, rendering
nodes for branching of diﬀerent amyloidogenic sequences(Figures 2I and S5).
We then performed LC-MS/MS analysis of peptides formed
after 1 min and 6 h of digestion at 2:1 excess of NE over Sprotein. We identiﬁed 98 NE cleavage peptides (Table S1) fromthe S-protein structure and classiﬁed these into three groups: (i)formed after 1 min (Figure 3A, B), (ii) formed after 1 min andstill present after 6 h (Figure 3A, C), and (iii) only present after 6
h (Figure 3A, D). Initial cleavage and further digestion (group i)
occurred mainly within the S2 domain with abundant cleavageof the HR domains and in the C-terminal part of S1. NTD and
RBD were much less aﬀected (Figure 3A). Three persistent
peptides formed after initial cleavage (group ii) originated from
NTD and RBD. Several peptides only formed after 6 h ofincubation (group iii). Strikingly, the peptide from segment194−203 (FKNIDGYFKI, included in Spike192) was part ofthis group and was highly abundant after 6 h (Table S2). Three
peptides containing segments from our seven spike peptides(Figure 3A, E) were formed as free peptides (Spike192,
Spike258, and Spike1166) still present after 6 h of co-incubation,
two were digested early and disappeared (Spike532 andSpike685), and two were likely still present in the parent nicked8947

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Figure 3. NE cleavage sites within full-length S-protein. Arrows indicate limits of the cryo-EM structure 27−1146. (A) Peptides identiﬁed by LC-MS/
MS (Table S1) in correlation to the amyloidogenic Spike peptides (cf. Figure 1 and Table 1) and the S-protein domain structure: after (i) 1 min (red),(ii) 1 min and still persistent at 6 h (magenta), (iii) only present after 6 h incubation (blue). (B−E) Same color code for cleaved peptide groups (i)−(iii) and spike peptides mapped onto the protomer cryo-EM S-protein structure PDB 6VXX.1

formation decreased the ﬁbrinolysis (Figure 4C). Furthermore,
the addition of 2% ﬁbrils (from 1 mg/mL stock, total 20 μg/mL)of Spike192 and 194−203 increased persistent plasmin
indigestible ﬁbrin (Figure 4 D). As expected, the more
amyloidogenic Spike192 induced more plasmin resistant ﬁbrinclots than did Spike194−203. Our reductionist assay appears to
replicate results from human plasma samples.7
We tested two ﬂuorescent analogues of positron emission
tomography (PET) amyloid tracers, CN-PiB (benzothiazole
analogue of Pittsburgh compound B) and DF-9 (stilbene
analogue of Florbetaben), known to bind to neurological Aβamyloid and cardiac AL, AA, and ATTR amyloid and foundstrong binding with concomitant ﬂuorescence response toward
Spike192 ﬁbrils in vitro (Figure S7). As a translational strategy,
PET imaging may hence serve as an option for clinical studies tocomplement liquid biopsies to assess amyloid microclots.26
In conclusion, we herein proposed a simple molecularmechanism for how SARS-CoV-2 S-protein endoproteolyzed
by NE can form amyloidogenic S-peptides, such as segment194−203, and lead to 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 if this process
occurs in vivo. We found that all common coronaviruses
infecting humans contain amyloidogenic sequences (Figure
S8A). Nonetheless, the magnitude of diverse COVID-19
symptoms was not previously reported. The segment 194−
213 is unique for SARS-CoV-2 (Figure S8B) which, in
combination with acute inﬂammation and neutrophil recruitment known to be more prevalent in COVID-19 compared toother viral infections, could explain the putative COVID-19

associated amyloid formation. It should be mentioned thatamyloidosis is rather common in the elderly population18 and its
associations with viral infections is a matter of discussion.13
Recent studies demonstrate that COVID-19 recovered patients
have an increased risk of type II diabetes, an amyloid associateddisease.28,29 While our study is limited to in vitro ﬁndings of pure
preparations of peptides and proteins, the results propose taking
S-protein amyloidogenesis into account when studying COVID19 and long COVID-19 symptoms.

■

ASSOCIATED CONTENT

sı Supporting Information*

The Supporting Information is available free of charge athttps://pubs.acs.org/doi/10.1021/jacs.2c03925.

■

Experimental procedures; bulk S-protein peptide librarysequences; selection of amyloidogenic segments for
peptide synthesis and comparison with peptides renderedby in silico elastase digestion (PDF)
Elastase peptides at 1 min vs 6 h (XLSX)Quantiﬁcation of abundant peaks (XLSX)AUTHOR INFORMATION

Corresponding Authors

Soﬁe Nyström − Department of Physics, Chemistry and Biology,
Linköping University, 58183 Linköping, Sweden;
orcid.org/0000-0002-4303-4783; Email: soﬁe.nystrom@liu.sePer Hammarström − Department of Physics, Chemistry andBiology, Linköping University, 58183 Linköping, Sweden;8948

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ThT, thioﬂavin T; CN-PiB, Cyano-Pittsburgh compound B;
TEM, transmission electron microscopy; DSF, diﬀerential
scanning ﬂuorimetry■

REFERENCES(1) Walls, A. C.; Park, Y. J.; Tortorici, M. A.; Wall, A.; McGuire, A. T.;
Veesler, D. Structure, Function, and Antigenicity of the SARS-CoV-2
Spike Glycoprotein. Cell 2020, 181 (2), 281−292.
(2) Huang, C.; Wang, Y.; Li, X.; Ren, L.; Zhao, J.; Hu, Y.; Zhang, L.;
Fan, G.; Xu, J.; Gu, X.; Cheng, Z.; Yu, T.; Xia, J.; Wei, Y.; Wu, W.; Xie,X.; Yin, W.; Li, H.; Liu, M.; Xiao, Y.; Gao, H.; Guo, L.; Xie, J.; Wang, G.;
Jiang, R.; Gao, Z.; Jin, Q.; Wang, J.; Cao, B. Clinical features of patients
infected with 2019 novel coronavirus in Wuhan, China. Lancet 2020,
395 (10223), 497−506.
(3) Lipcsey, M.; Persson, B.; Eriksson, O.; Blom, A. M.; Fromell, K.;
Hultstrom, M.; Huber-Lang, M.; Ekdahl, K. N.; Frithiof, R.; Nilsson, B.
The Outcome of Critically Ill COVID-19 Patients Is Linked toThromboinflammation Dominated by the Kallikrein/Kinin System.
Front Immunol 2021, 12, 627579.
(4) Gao, Y. M.; Xu, G.; Wang, B.; Liu, B. C. Cytokine storm syndromein coronavirus disease 2019: A narrative review. J. Intern Med. 2021, 289(2), 147−161.
(5) Sinha, N.; Thakur, A. K. Likelihood of amyloid formation in
COVID-19-induced ARDS. Trends Microbiol 2021, 29 (11), 967−969.
(6) Tavassoly, O.; Safavi, F.; Tavassoly, I. Seeding Brain Protein
Aggregation by SARS-CoV-2 as a Possible Long-Term Complication of
COVID-19 Infection. ACS Chem. Neurosci. 2020, 11 (22), 3704−3706.
(7) Grobbelaar, L. M.; Venter, C.; Vlok, M.; Ngoepe, M.; Laubscher,
G. J.; Lourens, P. J.; Steenkamp, J.; Kell, Douglas B.; Pretorius, E. SARSCoV-2 spike protein S1 induces fibrin(ogen) resistant to fibrinolysis:
implications for microclot formation in COVID-19. Biosci. Rep. 2021,
41 (8), BSR20210611.
(8) Bouma, B.; Maas, C.; Hazenberg, B. P.; Lokhorst, H. M.; Gebbink,
M. F. Increased plasmin-alpha2-antiplasmin levels indicate activation of
the fibrinolytic system in systemic amyloidoses. J. Thromb Haemost
2007, 5 (6), 1139−42.
(9) Hammarstrom, P. The bloody path of amyloids and prions. J.
Thromb Haemost 2007, 5 (6), 1136−8.
(10) Maas, C.; Govers-Riemslag, J. W.; Bouma, B.; Schiks, B.;
Hazenberg, B. P.; Lokhorst, H. M.; Hammarstrom, P.; ten Cate, H.; deGroot, P. G.; Bouma, B. N.; Gebbink, M. F. Misfolded proteins activate
factor XII in humans, leading to kallikrein formation without initiating
coagulation. J. Clin. Invest. 2008, 118 (9), 3208−18.
(11) Munch, J.; Rucker, E.; Standker, L.; Adermann, K.; Goffinet, C.;
Schindler, M.; Wildum, S.; Chinnadurai, R.; Rajan, D.; Specht, A.;
Gimenez-Gallego, G.; Sanchez, P. C.; Fowler, D. M.; Koulov, A.; Kelly,
J. W.; Mothes, W.; Grivel, J. C.; Margolis, L.; Keppler, O. T.;
Forssmann, W. G.; Kirchhoff, F. Semen-derived amyloid fibrils
drastically enhance HIV infection. Cell 2007, 131 (6), 1059−71.
(12) Tayeb-Fligelman, E.; Cheng, X.; Tai, C.; Bowler, J. T.; Griner, S.;
Sawaya, M. R.; Seidler, P. M.; Jiang, Y. X.; Lu, J.; Rosenberg, G. M.;
Salwinski, L.; Abskharon, R.; Zee, C. T.; Hou, K.; Li, Y.; Boyer, D. R.;
Murray, K. A.; Falcon, G.; Anderson, D. H.; Cascio, D.; Saelices, L.;
Damoiseaux, R.; Guo, F.; Eisenberg, D. S., Inhibition of amyloid
formation of the Nucleoprotein of SARS-CoV-2. bioRxiv, March 18,
2021, ver. 1. DOI: 10.1101/2021.03.05.434000v2 (accessed 2022-0309).
(13) Michiels, E.; Rousseau, F.; Schymkowitz, J. Mechanisms andtherapeutic potential of interactions between human amyloids andviruses. Cellular and molecular life sciences: CMLS 2021, 78 (6), 2485−
2501.
(14) Zhang, S. M.; Liao, Y.; Neo, T. L.; Lu, Y.; Liu, D. X.; Vahlne, A.;
Tam, J. P. Identification and application of self-binding zipper-likesequences in SARS-CoV spike protein. international journal ofbiochemistry & cell biology 2018, 101, 103−112.
(15) Idrees, D.; Kumar, V. SARS-CoV-2 spike protein interactions
with amyloidogenic proteins: Potential clues to neurodegeneration.
Biochemical and biophysical research communications 2021, 554, 94−98.

Figure 4. (A) Spike194−203 properties. (B) Spike194−203
amyloidogenicity monitored by ThT kinetics, Congo red birefringence,
and TEM. (C) Thrombin activated ﬁbrin formation and plasminogen/tPA-induced ﬁbrinolysis in the absence and presence of seven-peptidemix amyloid ﬁbrils (Table 1, Figure S3). (D) Turbidity diﬀerence afterﬁbrin formation and ﬁbrinolysis in the absence and presence of
Spike192 ﬁbrils and Spike194−203 ﬁbrils (n = 10−12 replicates of eachreaction, corrected for background scatter).

orcid.org/0000-0001-5827-3587;
Email: per.hammarstrom@liu.se
Complete contact information is available at:https://pubs.acs.org/10.1021/jacs.2c03925
Author Contributions†

The research was performed and the manuscript was writtenwith equal contributions of both authors.
Funding

This research was ﬁnanced by The Swedish Research Council
Grant #2019-04405.
Notes

The authors declare no competing ﬁnancial interest.

■

ACKNOWLEDGMENTS
We thank Björn Wallner for Alphafold 2 folding predictions;
Xiongyu Wu for synthesis of CN-PiB; Jun Zhang for synthesis of
DF-9; LiU core-facilities ProLinC; Medical Faculty Microscopy;
Proteomics core, especially Maria Turkina for collection ofexperimental data.

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ABBREVIATIONSCNS, central nervous system; SARS-CoV-2, Severe AcuteRespiratory Syndrome Coronavirus-2; S-protein, spike protein;8949

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pubs.acs.org/JACS(16) Maurer-Stroh, S.; Debulpaep, M.; Kuemmerer, N.; Lopez de laPaz, M.; Martins, I. C.; Reumers, J.; Morris, K. L.; Copland, A.; Serpell,
L.; Serrano, L.; Schymkowitz, J. W.; Rousseau, F. Exploring thesequence determinants of amyloid structure using position-specificscoring matrices. Nat. Methods 2010, 7 (3), 237−42.
(17) Upadhyay, V.; Lucas, A.; Panja, S.; Miyauchi, R.; Mallela, K. M.
G. Receptor binding, immune escape, and protein stability direct thenatural selection of SARS-CoV-2 variants. J. Biol. Chem. 2021, 297 (4),101208.
(18) Benson, M. D.; Buxbaum, J. N.; Eisenberg, D. S.; Merlini, G.;
Saraiva, M. J. M.; Sekijima, Y.; Sipe, J. D.; Westermark, P. Amyloid
nomenclature 2020: update and recommendations by the International
Society of Amyloidosis (ISA) nomenclature committee. Amyloid 2020,27 (4), 217−222.
(19) Sipe, J. D. Amyloid proteins: the beta sheet conformation anddisease; Wiley-VCH: Weinheim, 2005.
(20) Peacock, T. P.; Goldhill, D. H.; Zhou, J.; Baillon, L.; Frise, R.;
Swann, O. C.; Kugathasan, R.; Penn, R.; Brown, J. C.; Sanchez-David,
R. Y.; Braga, L.; Williamson, M. K.; Hassard, J. A.; Staller, E.; Hanley, B.;
Osborn, M.; Giacca, M.; Davidson, A. D.; Matthews, D. A.; Barclay, W.
S. The furin cleavage site in the SARS-CoV-2 spike protein is required
for transmission in ferrets. Nature microbiology 2021, 6 (7), 899−909.
(21) Johansson, C.; Kirsebom, F. C. M. Neutrophils in respiratory
viral infections. Mucosal immunology 2021, 14 (4), 815−827.
(22) Pandey, K. C.; De, S.; Mishra, P. K. Role of Proteases in Chronic
Obstructive Pulmonary Disease. Front. Pharmacol. 2017, 8, 512.
(23) Strnad, P.; McElvaney, N. G.; Lomas, D. A. Alpha1-Antitrypsin
Deficiency. N Engl J. Med. 2020, 382 (15), 1443−1455.
(24) Edwards, R. J.; Mansouri, K.; Stalls, V.; Manne, K.; Watts, B.;
Parks, R.; Janowska, K.; Gobeil, S. M. C.; Kopp, M.; Li, D.; Lu, X.; Mu,
Z.; Deyton, M.; Oguin, T. H., 3rd; Sprenz, J.; Williams, W.; Saunders, K.
O.; Montefiori, D.; Sempowski, G. D.; Henderson, R.; Munir Alam, S.;
Haynes, B. F.; Acharya, P. Cold sensitivity of the SARS-CoV-2 spikeectodomain. Nat. Struct Mol. Biol. 2021, 28 (2), 128−131.
(25) Mishra, R.; Sorgjerd, K.; Nystrom, S.; Nordigarden, A.; Yu, Y. C.;
Hammarstrom, P. Lysozyme amyloidogenesis is accelerated by specificnicking and fragmentation but decelerated by intact protein binding
and conversion. J. Mol. Biol. 2007, 366 (3), 1029−44.
(26) Kell, D. B.; Laubscher, G. J.; Pretorius, E. A central role foramyloid fibrin microclots in long COVID/PASC: origins andtherapeutic implications. Biochem. J. 2022, 479 (4), 537−559.
(27) Terasawa, F.; Kani, S.; Hongo, M.; Okumura, N. In vitro fibrin
clot formation and fibrinolysis using heterozygous plasma fibrinogenfrom gammaAsn319, Asp320 deletion dysfibrinogen, Otsu I. Thromb
Res. 2006, 118 (5), 651−61.
(28) Rathmann, W.; Kuss, O.; Kostev, K. Incidence of newly
diagnosed diabetes after Covid-19. Diabetologia 2022, 65, 949−954.
(29) Barrett, C. E.; Koyama, A. K.; Alvarez, P.; Chow, W.; Lundeen, E.
A.; Perrine, C. G.; Pavkov, M. E.; Rolka, D. B.; Wiltz, J. L.; BullOtterson, L.; Gray, S.; Boehmer, T. K.; Gundlapalli, A. V.; Siegel, D. A.;
Kompaniyets, L.; Goodman, A. B.; Mahon, B. E.; Tauxe, R. V.; Remley,
K.; Saydah, S. Risk for Newly Diagnosed Diabetes > 30 Days After
SARS-CoV-2 Infection Among Persons Aged < 18 Years - UnitedStates, March 1, 2020-June 28, 2021. MMWR Morb Mortal Wkly Rep
2022, 71 (2), 59−65.

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https://doi.org/10.1021/jacs.2c03925
J. Am. Chem. Soc. 2022, 144, 8945−8950