Membrane association and protein conformation of α-synuclein in intact neurons. Effect of Parkinson's disease-linked mutations

Pamela J McLean, Hibiki Kawamata, Scott Ribich, Bradley T. Hyman

Research output: Contribution to journalArticle

201 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)8812-8816
Number of pages5
JournalJournal of Biological Chemistry
Volume275
Issue number12
DOIs
StatePublished - Mar 24 2000
Externally publishedYes

Fingerprint

Synucleins
Protein Conformation
Neurons
Parkinson Disease
Conformations
Membrane Proteins
Association reactions
Membranes
Mutation
Proteins
Fluorescence Resonance Energy Transfer
Lewy Bodies
Lewy Body Disease
Gene encoding
Missense Mutation
Transfection

ASJC Scopus subject areas

  • Biochemistry

Cite this

Membrane association and protein conformation of α-synuclein in intact neurons. Effect of Parkinson's disease-linked mutations. / McLean, Pamela J; Kawamata, Hibiki; Ribich, Scott; Hyman, Bradley T.

In: Journal of Biological Chemistry, Vol. 275, No. 12, 24.03.2000, p. 8812-8816.

Research output: Contribution to journalArticle

@article{6f3d68f109c64645a7f57bb031695166,
title = "Membrane association and protein conformation of α-synuclein in intact neurons. Effect of Parkinson's disease-linked mutations",
abstract = "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.",
author = "McLean, {Pamela J} and Hibiki Kawamata and Scott Ribich and Hyman, {Bradley T.}",
year = "2000",
month = "3",
day = "24",
doi = "10.1074/jbc.275.12.8812",
language = "English (US)",
volume = "275",
pages = "8812--8816",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "12",

}

TY - JOUR

T1 - Membrane association and protein conformation of α-synuclein in intact neurons. Effect of Parkinson's disease-linked mutations

AU - McLean, Pamela J

AU - Kawamata, Hibiki

AU - Ribich, Scott

AU - Hyman, Bradley T.

PY - 2000/3/24

Y1 - 2000/3/24

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0034708767&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034708767&partnerID=8YFLogxK

U2 - 10.1074/jbc.275.12.8812

DO - 10.1074/jbc.275.12.8812

M3 - Article

C2 - 10722726

AN - SCOPUS:0034708767

VL - 275

SP - 8812

EP - 8816

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 12

ER -