Interaction of alpha-synuclein and synphilin-1: effect of Parkinson's disease-associated mutations.
alpha-Synuclein is a major component of Lewy bodies, a neuropathological feature of Parkinson's disease. Two alpha-synuclein mutations, Ala53Thr and Ala30Pro, are associated with early onset, familial forms of the disease. Recently, synphilin-1, a protein found to interact with alpha-synuclein by yeast two hybrid techniques, was detected in Lewy bodies. In this study we report the interaction of alpha-synuclein and synphilin-1 in human neuroglioma cells using a sensitive fluorescence resonance energy transfer technique. We demonstrate that the C-terminus of alpha-synuclein is closely associated with the C-terminus of synphilin-1. A weak interaction occurs between the N-terminus of alpha-synuclein and synphilin-1. The familial Parkinson's disease associated mutations of alpha-synuclein (Ala53Thr and Ala30Pro) also demonstrate a strong interaction between their C-terminal regions and synphilin-1. However, compared with wild-type alpha-synuclein, significantly less energy transfer occurs between the C-terminus of Ala53Thr alpha-synuclein and synphilin-1, suggesting that the Ala53Thr mutation alters the conformation of alpha-synuclein in relation to synphilin-1.
Kawamata
H
H
Alzheimer's Disease Research Unit, Department of Neurology, Massachusetts General Hospital East, Charlestown 02129, USA.
McLean
P J
PJ
Sharma
N
N
Hyman
B T
BT
D015153:Blotting, Western; D002352:Carrier Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D004735:Energy Transfer; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D005910:Glioma; D006801:Humans; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010300:Parkinson Disease / Q000235:genetics*; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013347:Subcellular Fractions / Q000737:chemistry; D051843:Synucleins; D014162:Transfection; D014407:Tumor Cells, Cultured; D051844:alpha-Synuclein
D002352:Carrier Proteins; D009419:Nerve Tissue Proteins; C497604:SNCA protein, human; C118958:SNCAIP protein, human; D051843:Synucleins; D051844:alpha-Synuclein