Helix periodicity, topology, and dynamics of membrane-associated alpha-synuclein.
The protein alpha-Synuclein (aS) is a synaptic vesicle-associated regulator of synaptic strength and dopamine homeostasis with a pathological role in Parkinson's disease. The normal function of aS depends on a membrane-associated conformation that is adopted upon binding to negatively charged lipid surfaces. Previously we found that the membrane-binding domain of aS is helical and suggested that it may exhibit an unusual structural periodicity. Here we present a study of the periodicity, topology, and dynamics of detergent micelle-bound aS using paramagnetic spin labels embedded in the micelle or attached to the protein. We show that the helical region of aS completes three full turns every 11 residues, demonstrating the proposed 11/3 periodicity. We also find that the membrane-binding domain is partially buried in the micelle surface and bends toward the hydrophobic interior, but does not traverse the micelle. Deeper submersion of certain regions within the micelle, including the unique lysine-free sixth 11-residue repeat, is observed and may be functionally important. There are no long-range tertiary contacts within this domain, indicating a highly extended configuration. The backbone dynamics of the micelle-bound region are relatively uniform with a slight decrease in flexibility observed toward the C-terminal end. These results clarify the topological features of aS bound to membrane-mimicking detergent micelles, with implications for aS function and pathology.
Bussell
Robert
R
Department of Biochemistry, Weill Medical College of Cornell University, 1300 York Avenue, New York, NY 10021, USA.
Ramlall
Trudy Fiona
TF
Eliezer
David
D
D000595:Amino Acid Sequence; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry*; D008670:Metals / Q000737:chemistry; D008823:Micelles; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D012997:Solvents / Q000737:chemistry; D013113:Spin Labels; D051843:Synucleins; D051844:alpha-Synuclein
D008565:Membrane Proteins; D008670:Metals; D008823:Micelles; D009419:Nerve Tissue Proteins; C497604:SNCA protein, human; D012997:Solvents; D013113:Spin Labels; D051843:Synucleins; D051844:alpha-Synuclein
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