Copper binding regulates intracellular alpha-synuclein localisation, aggregation and toxicity.
Alpha-synuclein is a natively unfolded protein that aggregates and forms inclusions that are associated with a range of diseases that include Parkinson's Disease and Dementia with Lewy Bodies. The mechanism behind the formation of these inclusions and their possible role in disease remains unclear. Alpha-synuclein has also been shown to bind metals including copper and iron. We used a cell culture model of alpha-synuclein aggregation to examine the relationship between metals and formation of aggregates of the protein. While the levels of iron appear to have no role in aggregate formation or localisation of the protein in cells, copper appears to be important for both aggregation and cellular localisation of alpha-synuclein. Reduction in cellular copper resulted in a great decrease in aggregate formation both in terms of large aggregates visible in cells and oligomers observed in western blot analysis of cell extracts. Reduction in copper also resulted in a change in localisation of the protein which became more intensely localised to the plasma membrane in medium with low copper. These changes were reversed when copper was restored to the cells. Mutants of the copper binding domains altered the response to copper. Deletion of either the N- or C-termini resulted in a loss of aggregation while deletion of the C-termini also resulted in a loss of membrane association. Increased expression of alpha-synuclein also increased cell sensitivity to the toxicity of copper. These results suggest that the potential pathological role of alpha-synuclein aggregates is dependent upon the copper binding capacity of the protein.
Department of Biology and Biochemistry, University of Bath, Bath, UK.
Xiaoyan
X
Wang
Dima
D
Moualla
Josephine A
JA
Wright
David R
DR
Brown
D001665:Binding Sites / Q000187:drug effects; D015153:Blotting, Western; D002449:Cell Aggregation / Q000187:drug effects; D016923:Cell Death / Q000502:physiology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D003001:Cloning, Molecular; D003300:Copper / Q000378:metabolism* / Q000502:physiology*; D006801:Humans; D018613:Microscopy, Confocal; D016296:Mutagenesis; D017354:Point Mutation / Q000502:physiology; D014162:Transfection; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity*
D051844:alpha-Synuclein; D003300:Copper