MOLECULAR BASIS OF CELL AND DEVELOPMENTAL BIOLOGY
Membrane Association and Protein Conformation of α-Synuclein in Intact Neurons: EFFECT OF PARKINSON′S DISEASE-LINKED MUTATIONS*

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Two missense mutations (Ala-30 → Pro and Ala-53 → Thr) in the gene encoding α-synuclein are associated with rare autosomal dominant forms of familial Parkinson's disease. In addition, α-synuclein is an abundant component of Lewy bodies in sporadic Parkinson's disease and diffuse Lewy body disease. However, the normal conformation of α-synuclein, its cellular localization in neurons, and the effects of the mutations remain to be determined. In the present study, we examine these questions using sensitive fluorescence resonance energy transfer techniques. Transient transfection of α-synuclein expression constructs into primary cortical neurons and counterstaining with the lipophilic fluorescent marker, DiI, demonstrates a close association between α-synuclein and cellular membranes. Both the N- and C-terminal regions of α-synuclein are tightly associated with membranes. A weak interaction also occurs between the N and C termini themselves. The Parkinson's disease-associated mutations have no effect on membrane interaction; however, the Ala-30 → Pro mutation alters the three-dimensional conformation of α-synuclein, as measured by significantly increased fluorescence resonance energy transfer between the N and C termini.

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This work was supported in part by Massachusetts General Hospital-Massachusetts Institute of Technology Parkinson's Disease Research Center Grant PONS 38372.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Recipient of an Massachusetts Biomedical Research Corp. Tosteson postdoctoral fellowship award.