Amyloid-Like Aggregation in Native Protein and its Suppression in the Bio-Conjugated Counterpart

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cam.issuedOnline2022-06-20
dc.contributor.authorMukhopadhyay, A
dc.contributor.authorStoev, ID
dc.contributor.authorKing, DA
dc.contributor.authorSharma, KP
dc.contributor.authorEiser, E
dc.date.accessioned2022-07-04T15:00:33Z
dc.date.available2022-07-04T15:00:33Z
dc.date.issued2022
dc.date.submitted2022-04-20
dc.date.updated2022-07-04T15:00:32Z
dc.description.abstract<jats:p>Prevention of protein aggregation and thus stabilization of proteins has large biological and biotechnological implications. Here we introduce Dynamic Light Scattering (DLS) and DLS-based microrheology to show how native bovine serum albumin (nBSA) forms amyloid fibrils in weakly denaturing conditions as function of time, and how stoichiometric conjugation of BSA with polymer-surfactants (PSpBSA) protects the protein form such aggregation. Employing a combination of Thioflavin-T fluorescence, Fourier transform infrared spectroscopy and other methods, we show that nBSA forms filamentous aggregates with amyloid-like structure, while PSpBSA proteins remain fully dispersed with only minor changes in their folding state, even when continuously heated for up to 5 days in denaturation conditions at 65 °C. Time-resolved DLS-based microrheology studies demonstrate that suspensions of the filamentous nBSA aggregates become viscoelastic for concentrations ≥200 <jats:italic>μ</jats:italic>M. Our results indicate that after 6 days in aggregation conditions, the elastic modulus <jats:italic>G</jats:italic>′(<jats:italic>ω</jats:italic>) of nBSA solutions went from zero initially to values of up to 3.6 Pa, indicating that the filaments become long enough to form an entangled, viscoelastic network. Interestingly, heating 200 <jats:italic>μ</jats:italic>M native BSA solutions at 65 °C for 2 days in Eppendorf tubes resulted in self-standing films rather than dispersed filaments. These films exhibited strong ThT-fluorescence intensities and a predominant <jats:italic>β</jats:italic>-sheet secondary structure in FTIR studies, suggesting that the self-standing microstructure of the film resulted from hierarchical self-assembly of the amyloid fibrils.</jats:p>
dc.identifier.citationFrontiers in Physics, volume 10, article-number 924864
dc.identifier.doi10.17863/CAM.86147
dc.identifier.eissn2296-424X
dc.identifier.issn2296-424X
dc.identifier.other924864
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/338736
dc.languageen
dc.publisherFrontiers Media SA
dc.publisher.urlhttp://dx.doi.org/10.3389/fphy.2022.924864
dc.subjectprotein aggregation
dc.subjectnBSA
dc.subjectamyloid fibrils
dc.subjectbio-conjugated protein
dc.subjectdynamic light scattering
dc.subjectmicrorheology
dc.subjectstability against denaturation
dc.titleAmyloid-Like Aggregation in Native Protein and its Suppression in the Bio-Conjugated Counterpart
dc.typeArticle
dcterms.dateAccepted2022-05-19
prism.publicationNameFrontiers in Physics
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/
rioxxterms.versionVoR
rioxxterms.versionofrecord10.3389/fphy.2022.924864
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