Defective membrane interactions of familial Parkinson's disease mutant A30P alpha-synuclein.
alpha-Synuclein (alpha-Syn) is an abundant presynaptic protein of unknown function, which has been implicated in the pathogenesis of Parkinson's disease. Alpha-Syn has been suggested to play a role in lipid transport and synaptogenesis, and growing evidence suggests that alpha-Syn interactions with cellular membranes are physiologically important. In the current study, we demonstrate that the familial Parkinson's disease-linked A30P mutant alpha-Syn is defective in binding to phospholipid vesicles in vitro as determined by vesicle ultracentrifugation, circular dichroism spectroscopy, and low-angle X-ray diffraction. Interestingly, our data also suggest that alpha-Syn may bind to the lipid vesicles as a dimer, which suggest that this species could be a physiologically relevant and functional entity. In contrast, the naturally occurring murine A53T substitution, which is also linked to Parkinson's disease, displayed a normal membrane-binding activity that was comparable to wild-type alpha-Syn. A double mutant A53T/A30P alpha-Syn showed defective membrane binding similar to the A30P protein, indicating that the proline mutation is dominant in terms of impairing the membrane-binding activity. With these observations, we suggest that the A53T and A30P mutants may have different physiological consequences in vivo and could possibly contribute to early onset Parkinson's disease via unique mechanisms.
Jo
Euijung
E
Centre for Research in Neurodegenerative Diseases, 6 Queen's Park Crescent West, Toronto, Ontario, M5S 3H2, Canada. euijung.jo@utoronto.ca
Fuller
Nola
N
Rand
R Peter
RP
St George-Hyslop
Peter
P
Fraser
Paul E
PE
D015153:Blotting, Western; D002942:Circular Dichroism; D019281:Dimerization; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020125:Mutation, Missense / Q000235:genetics*; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D009994:Osmolar Concentration; D009997:Osmotic Pressure; D010300:Parkinson Disease / Q000235:genetics*; D010743:Phospholipids / Q000378:metabolism; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D055672:Static Electricity; D051843:Synucleins; D014461:Ultracentrifugation; D014961:X-Ray Diffraction; D051844:alpha-Synuclein
D008051:Lipid Bilayers; D008081:Liposomes; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D010743:Phospholipids; C497604:SNCA protein, human; D051843:Synucleins; D051844:alpha-Synuclein