title: The E46K mutation in alpha-synuclein increases amyloid fibril formation.
abstract: The identification of a novel mutation (E46K) in one of the KTKEGV-type repeats in the amino-terminal region of alpha-synuclein suggests that this region and, more specifically, Glu residues in the repeats may be important in regulating the ability of alpha-synuclein to polymerize into amyloid fibrils. It was demonstrated that the E46K mutation increased the propensity of alpha-synuclein to fibrillize, but this effect was less than that of the A53T mutation. The substitution of Glu(46) for an Ala also increased the assembly of alpha-synuclein, but the polymers formed can have different ultrastructures, further indicating that this amino acid position has a significant effect on the assembly process. The effect of residue Glu(83) in the sixth repeat of alpha-synuclein, which lies closest to the amino acid stretch critical for filament assembly, was also studied. Mutation of Glu(83) to a Lys or Ala increased polymerization but perturbed some of the properties of mature amyloid. These results demonstrated that some of the Glu residues within the repeats can have significant effects on modulating the assembly of alpha-synuclein to form amyloid fibrils. The greater effect of the A53T mutation, even when compared with what may be predicted to be a more dramatic mutation such as E46K, underscores the importance of protein microenvironment in affecting protein structure. Moreover, the relative effects of the A53T and E46K mutations are consistent with the age of onset of disease. These findings support the notion that aberrant alpha-synuclein polymerization resulting in the formation of pathological inclusions can lead to disease.
authors.affiliation: Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA.
authors.forename: Eric A
authors.initials: EA
authors.lastname: Greenbaum
authors.forename: Charles L
authors.initials: CL
authors.lastname: Graves
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Mishizen-Eberz
authors.forename: Michael A
authors.initials: MA
authors.lastname: Lupoli
authors.forename: David R
authors.initials: DR
authors.lastname: Lynch
authors.forename: S Walter
authors.initials: SW
authors.lastname: Englander
authors.forename: Paul H
authors.initials: PH
authors.lastname: Axelsen
authors.forename: Benoit I
authors.initials: BI
authors.lastname: Giasson
mesh_terms: D000409:Alanine / Q000737:chemistry; D000595:Amino Acid Sequence; D000682:Amyloid / Q000737:chemistry*; D015153:Blotting, Western; D002154:Calpain / Q000737:chemistry; D002850:Chromatography, Gel; D002942:Circular Dichroism; D018076:DNA, Complementary / Q000378:metabolism; D018698:Glutamic Acid / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008239:Lysine / Q000737:chemistry; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics*; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D017550:Spectroscopy, Fourier Transform Infrared; D051843:Synucleins; D013997:Time Factors; D051844:alpha-Synuclein
chemical_list: D000682:Amyloid; D018076:DNA, Complementary; D009419:Nerve Tissue Proteins; C497604:SNCA protein, human; D051843:Synucleins; D051844:alpha-Synuclein; D018698:Glutamic Acid; D002154:Calpain; D008239:Lysine; D000409:Alanine