The impact of the E46K mutation on the properties of α-synuclein in its monomelic and oligomeric states

Ross A. Fredenburg, Carla Rospigliosi, Robin K. Meray, Jeffrey C. Kessler, Hilal A. Lashuel, David Eliezer, Peter T. Lansbury

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Abstract

The third and most recently identified Parkinson's disease-linked variant of the neuronal protein α-synuclein to be identified (E46K) results in widespread brain pathology and early onset Parkinson symptoms (Zarranz et al. (2004) Ann. Neurol. 55, 164-173). Herein, we present biochemical and biophysical characterization of E46K α-synuclein in various states of aggregation. Circular dichroism and nuclear magnetic resonance spectroscopy illustrate that the E46K mutation results in subtle changes in the conformation of the monomeric protein both free in solution and in the presence of SDS micelles. However, it does not alter the overall helical propensity of the protein in the presence of phospholipids. E46K α-synuclein formed insoluble fibrils in vitro more rapidly than the wild type protein, and electron microscopy revealed that E46K α-synuclein fibrils possess a typical amyloid ultrastructure. E46K α-synuclein protofibrils, soluble aggregates that form during the transition from the monomeric form to the fibrillar form of α-synuclein, were characterized by electron microscopy and gel filtration and were found to include annular species. The unique ability of a subfraction of E46K and wild type α-synuclein protofibrils containing porelike species to permeabilize lipid vesicles was demonstrated in vitro using a real-time chromatographic method. In contrast to simplistic expectations, the total amount of protofibrils and the amount of permeabilizing activity per mole protein in the protofibril fraction were reduced by the E46K mutation. These results suggest that if the porelike activity of α-synuclein is important for neurotoxicity, there must be factors in the neuronal cytoplasm that reverse the trends in the intrinsic properties of E46K versus WT α-synuclein that are observed in vitro.

Original languageEnglish
Pages (from-to)7107-7118
Number of pages12
JournalBiochemistry
Volume46
Issue number24
DOIs
Publication statusPublished - 19 Jun 2007
Externally publishedYes

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Synucleins
Mutation
Proteins
Electron microscopy
Electron Microscopy
Protein Conformation
Micelles
Pathology
Circular Dichroism
Amyloid
Nuclear magnetic resonance spectroscopy
Gel Chromatography
Parkinson Disease
Conformations
Brain
Phospholipids
Cytoplasm
Magnetic Resonance Spectroscopy
Agglomeration
Gels

ASJC Scopus subject areas

  • Biochemistry

Cite this

Fredenburg, R. A., Rospigliosi, C., Meray, R. K., Kessler, J. C., Lashuel, H. A., Eliezer, D., & Lansbury, P. T. (2007). The impact of the E46K mutation on the properties of α-synuclein in its monomelic and oligomeric states. Biochemistry, 46(24), 7107-7118. https://doi.org/10.1021/bi7000246

The impact of the E46K mutation on the properties of α-synuclein in its monomelic and oligomeric states. / Fredenburg, Ross A.; Rospigliosi, Carla; Meray, Robin K.; Kessler, Jeffrey C.; Lashuel, Hilal A.; Eliezer, David; Lansbury, Peter T.

In: Biochemistry, Vol. 46, No. 24, 19.06.2007, p. 7107-7118.

Research output: Contribution to journalArticle

Fredenburg, RA, Rospigliosi, C, Meray, RK, Kessler, JC, Lashuel, HA, Eliezer, D & Lansbury, PT 2007, 'The impact of the E46K mutation on the properties of α-synuclein in its monomelic and oligomeric states', Biochemistry, vol. 46, no. 24, pp. 7107-7118. https://doi.org/10.1021/bi7000246
Fredenburg RA, Rospigliosi C, Meray RK, Kessler JC, Lashuel HA, Eliezer D et al. The impact of the E46K mutation on the properties of α-synuclein in its monomelic and oligomeric states. Biochemistry. 2007 Jun 19;46(24):7107-7118. https://doi.org/10.1021/bi7000246
Fredenburg, Ross A. ; Rospigliosi, Carla ; Meray, Robin K. ; Kessler, Jeffrey C. ; Lashuel, Hilal A. ; Eliezer, David ; Lansbury, Peter T. / The impact of the E46K mutation on the properties of α-synuclein in its monomelic and oligomeric states. In: Biochemistry. 2007 ; Vol. 46, No. 24. pp. 7107-7118.
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