Effects of Parkinson's disease-linked mutations on the structure of lipid-associated alpha-synuclein.
Alpha-synuclein (alphaS) is a lipid-binding synaptic protein of unknown function that is found in an aggregated amyloid fibril form in the intraneuronal Lewy body deposits that are a defining characteristic of Parkinson's disease (PD). Although intrinsically unstructured when free in solution, alphaS adopts a highly helical conformation in association with lipid membranes or membrane mimetic detergent micelles. Two mutations in the alphaS gene have been linked to early onset autosomal dominant hereditary forms of PD, and have been shown to affect the aggregation kinetics of the protein in vitro. We have used high-resolution NMR spectroscopy, circular dichroism, and limited proteolysis to investigate the effects of these PD-linked mutations on the helical structure adopted by alphaS in the lipid or detergent micelle-bound form. We show that neither the A53T nor the A30P mutation has a significant effect on the structure of the folded protein, although the A30P mutation may cause a minor perturbation in the helical structure around the site of the mutation. The A30P, but not the A53T, mutation also appears to decrease the affinity of the protein for lipid surfaces, possibly by perturbing the nascent helical structure of the free protein. The potential implications of these results for the role of alphaS in PD are discussed.
Bussell
Robert
R
Department of Physiology, Biophysics and Molecular Medicine, Weill Medical College of Cornell University, 1300 York Avenue, New York, New York 10021, USA.
Eliezer
David
D
D000409:Alanine / Q000235:genetics; D006801:Humans; D050356:Lipid Metabolism; D008081:Liposomes; D008823:Micelles; D020125:Mutation, Missense; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular; D010300:Parkinson Disease / Q000235:genetics*; D010712:Phosphatidic Acids / Q000378:metabolism; D010713:Phosphatidylcholines / Q000378:metabolism; D011392:Proline / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D017433:Protein Structure, Secondary / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051843:Synucleins; D013912:Threonine / Q000235:genetics; D051844:alpha-Synuclein
C103238:1-palmitoyl-2-oleoyl-glycero-3-phosphatidic acid; D008081:Liposomes; D008823:Micelles; D009419:Nerve Tissue Proteins; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D011994:Recombinant Proteins; C497604:SNCA protein, human; D051843:Synucleins; D051844:alpha-Synuclein; D013912:Threonine; D011392:Proline; D000409:Alanine; C028694:1-palmitoyl-2-oleoylphosphatidylcholine