Publication year
2014Source
Biochimica et Biophysica Acta. Proteins and Proteomics, 1844, 12, (2014), pp. 2127-2134ISSN
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Publication type
Article / Letter to editor
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Organization
Tumorimmunology
Journal title
Biochimica et Biophysica Acta. Proteins and Proteomics
Volume
vol. 1844
Issue
iss. 12
Page start
p. 2127
Page end
p. 2134
Subject
Radboudumc 2: Cancer development and immune defence RIMLS: Radboud Institute for Molecular Life SciencesAbstract
The intrinsically disordered human alpha-synuclein (alphaSyn) protein exhibits considerable heterogeneity in in vitro fibrillization reactions. Using atomic force microscopy (AFM) we show that depending on the solvent conditions, A140C mutant and wild-type alphaSyn can be directed to reproducibly form homogeneous populations of fibrils exhibiting regular periodicity. Results from Thioflavin-T fluorescence assays, determination of residual monomer concentrations and native polyacrylamide gel electrophoresis reveal that solvent conditions including EDTA facilitate incorporation of a larger fraction of monomers into fibrils. The fibrils formed in 10mM Tris-HCl, 10mM NaCl and 0.1mM EDTA at pH7.4 display a narrow distribution of periodicities with an average value of 102+/-6nm for the A140C mutant and 107+/-9nm for wt alphaSyn. The ability to produce a homogeneous fibril population can be instrumental in understanding the detailed structural features of fibrils and the fibril assembly process. Moreover, the availability of morphologically well-defined fibrils will enhance the potential for use of amyloids as biological nanomaterials.
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- Faculty of Medical Sciences [93209]
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