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Tracking the heterogeneous distribution of amyloid spherulites and their population balance with free fibrils
- Source :
- The European physical journal. E, Soft matter. 33(4)
- Publication Year :
- 2010
-
Abstract
- The analysis of amyloidogenic systems reveals the appearance of distinct states of aggregation for amyloid fibrils. For different proteins and under specific experimental conditions, amyloid spherulites are recognized as a significant component occurring in several protein model systems used for in vitro fibrillation studies. In this work we have developed an approach to characterize solutions containing a mixture of amyloid spherulites and individual fibrils. Using bovine insulin as the model system, sedimentation kinetics for the amyloid aggregates were followed using a combination of UV-Vis spectroscopy and cross-polarized optical microscopy. Spherulites were identified as the species undergoing sedimentation. A simple mathematical approach allows the description of the kinetics in terms of decay time/rate distribution. Moreover, based on the sedimentation kinetics, a rough estimate of the balance between amyloid spherulites and individual fibrils can be provided. Fitting the experimental data with the proposed physico-chemical approach shows self-consistent results in reasonable agreement with quantitative imaging analysis previously reported. Our results provide new physical insights into the analysis of amyloidogenic systems, providing a method to characterize the heterogeneous distribution of amyloid spherulites and simultaneously distinguish spherulites and free fibril populations. Importantly, the method can be generally applied to the characterization of polydisperse solutions containing optically traceable spherical particles in the micrometric range.
- Subjects :
- Amyloid
Protein Folding
Materials science
Protein Conformation
Kinetics
Population
Biophysics
Nanotechnology
Fibril
Biochemistry
Protein structure
Microscopy
Animals
Insulin
General Materials Science
Soft matter
education
education.field_of_study
Surfaces and Interfaces
General Chemistry
Solutions
Models, Chemical
Protein folding
Cattle
Spectrophotometry, Ultraviolet
Microscopy, Polarization
Biotechnology
Protein Binding
Subjects
Details
- ISSN :
- 1292895X
- Volume :
- 33
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- The European physical journal. E, Soft matter
- Accession number :
- edsair.doi.dedup.....1c4d28ce02140e3f93861ed3f0077724