000056279 001__ 56279
000056279 005__ 20200221144324.0
000056279 0247_ $$2doi$$a10.1073/pnas.1524128113
000056279 0248_ $$2sideral$$a94043
000056279 037__ $$aART-2016-94043
000056279 041__ $$aeng
000056279 100__ $$aIljina, M.
000056279 245__ $$aKinetic model of the aggregation of alpha-synuclein provides insights into prion-like spreading
000056279 260__ $$c2016
000056279 5060_ $$aAccess copy available to the general public$$fUnrestricted
000056279 5203_ $$aThe protein alpha-synuclein (aS) self-assembles into small oligomeric species and subsequently into amyloid fibrils that accumulate and proliferate during the development of Parkinson''s disease. However, the quantitative characterization of the aggregation and spreading of aS remains challenging to achieve. Previously, we identified a conformational conversion step leading from the initially formed oligomers to more compact oligomers preceding fibril formation. Here, by a combination of single-molecule fluorescencemeasurements and kinetic analysis, we find that the reaction in solution involves two unimolecular structural conversion steps, from the disordered to more compact oligomers and then to fibrils, which can elongate by further monomer addition. We have obtained individual rate constants for these key microscopic steps by applying a global kinetic analysis to both the decrease in the concentration of monomeric protein molecules and the increase in oligomer concentrations over a 0.5-140-µM range of aS. The resulting explicit kinetic model of aS aggregation has been used to quantitatively explore seeding the reaction by either the compact oligomers or fibrils. Our predictions reveal that, although fibrils are more effective at seeding than oligomers, very high numbers of seeds of either type, of the order of 104, are required to achieve efficient seeding and bypass the slow generation of aggregates through primary nucleation. Complementary cellular experiments demonstrated that two orders of magnitude lower numbers of oligomers were sufficient to generate high levels of reactive oxygen species, suggesting that effective templated seeding is likely to require both the presence of template aggregates and conditions of cellular stress.
000056279 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000056279 590__ $$a9.661$$b2016
000056279 591__ $$aMULTIDISCIPLINARY SCIENCES$$b4 / 63 = 0.063$$c2016$$dQ1$$eT1
000056279 592__ $$a6.575$$b2016
000056279 593__ $$aMultidisciplinary$$c2016$$dQ1
000056279 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000056279 700__ $$aGarcia, G.A.
000056279 700__ $$aHorrocks, M.H.
000056279 700__ $$aTosatto, L.
000056279 700__ $$aChoi, M.L.
000056279 700__ $$aGanzinger, K.A.
000056279 700__ $$aAbramov, A.Y.
000056279 700__ $$aGandhi, S.
000056279 700__ $$aWood, N.W.
000056279 700__ $$0(orcid)0000-0002-9138-6687$$aCremades, N.$$uUniversidad de Zaragoza
000056279 700__ $$aDobson, C.M.
000056279 700__ $$aKnowles, T.P.J.
000056279 700__ $$aKlenerman, D.
000056279 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000056279 773__ $$g113, 9 (2016), E1206-E1215$$pProc. Natl. Acad. Sci.$$tProceedings of the National Academy of Sciences$$x0027-8424
000056279 8564_ $$s1628280$$uhttps://zaguan.unizar.es/record/56279/files/texto_completo.pdf$$yVersión publicada
000056279 8564_ $$s142702$$uhttps://zaguan.unizar.es/record/56279/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000056279 909CO $$ooai:zaguan.unizar.es:56279$$particulos$$pdriver
000056279 951__ $$a2020-02-21-13:42:58
000056279 980__ $$aARTICLE