RT Journal Article
T1 Probing the Occurrence of Soluble Oligomers through Amyloid Aggregation Scaling Laws
A1 Silva, Alexandra
A1 Sárkány, Zsuzsa
A1 Fraga, Joana S.
A1 Taboada Antelo, Pablo
A1 Ribeiro, Sandra Macedo
A1 Martins, Pedro M.
K1 Protein aggregation
K1 Amyloid
K1 Soluble oligomers
K1 Kinetic analysis
K1 Nucleation
AB Drug discovery frequently relies on the kinetic analysis of physicochemical reactions thatare at the origin of the disease state. Amyloid fibril formation has been extensively investigated inrelation to prevalent and rare neurodegenerative diseases, but thus far no therapeutic solution hasdirectly arisen from this knowledge. Other aggregation pathways producing smaller, hard-to-detectsoluble oligomers are increasingly appointed as the main reason for cell toxicity and cell-to-celltransmissibility. Here we show that amyloid fibrillation kinetics can be used to unveil the proteinoligomerization state. This is illustrated for human insulin and ataxin-3, two model proteins for whichthe amyloidogenic and oligomeric pathways are well characterized. Aggregation curves measuredby the standard thioflavin-T (ThT) fluorescence assay are shown to reflect the relative compositionof protein monomers and soluble oligomers measured by nuclear magnetic resonance (NMR) forhuman insulin, and by dynamic light scattering (DLS) for ataxin-3. Unconventional scaling laws ofkinetic measurables were explained using a single set of model parameters consisting of two rateconstants, and in the case of ataxin-3, an additional order-of-reaction. The same fitted parameterswere used in a discretized population balance that adequately describes time-course measurementsof fibril size distributions. Our results provide the opportunity to study oligomeric targets usingsimple, high-throughput compatible, biophysical assays.
PB MDPI
YR 2018
FD 2018
LK http://hdl.handle.net/10347/22804
UL http://hdl.handle.net/10347/22804
LA eng
NO Silva, A.; Sárkány, Z.; Fraga, J.S.; Taboada, P.; Macedo-Ribeiro, S.; Martins, P.M. Probing the Occurrence of Soluble Oligomers through Amyloid Aggregation Scaling Laws. Biomolecules 2018, 8, 108
NO This work was financed by (i) FEDER—Fundo Europeu de Desenvolvimento Regional funds through theCOMPETE 2020—Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020,and by Portuguese funds through FCT—Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologiae Ensino Superior in the framework of the projects POCI-01-0145-FEDER-031173 (PTDC/BIA-BFS/31173/2017)and POCI-01-0145-FEDER-007274 (“Institute for Research and Innovation in Health Sciences”), and by (ii)FEDER through Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020Partnership Agreement in the framework of Project Norte-01-0145-FEDER-000008. A.S. thanks the AmyloidosisFoundation (USA). P.T. thanks Ministerio de Economía y Competitividad (MINECO) and FEDER for researchproject MAT 2016-80266-R
DS Minerva
RD 29 abr 2026