title: In vitro formation of amyloid from alpha-synuclein is dominated by reactions at hydrophobic interfaces.
abstract: Most in vitro investigations of alpha-Synuclein (alphaSyn) aggregation and amyloidogenesis use agitation in the presence of air and/or Teflon to accelerate kinetics. The effect of the agitation is implicitly or explicitly attributed to mass transfer or fibril fragmentation. This paper evaluates these hypotheses by agitating alphaSyn under typical amyloidogenic conditions with controlled numbers of balls made of polytetrafluoroethylene (PTFE), polymethylmethacrylate (PMMA), and borosilicate glass with no headspace. Amyloid was assayed using thioflavin T fluorescence and atomic force microscopy. The observed kinetics were proportional to the PTFE surface area; the effects of PMMA and glass balls were negligible by comparison. No amyloid was observed to form in the absence of mixing balls. Agitation with only air also showed accelerated kinetics but different aggregate morphology. The results indicate that the mechanism active in agitation experiments is dominated by reactions at the hydrophobic-water interface. Of the mass transfer, fragmentation, and hydrophobic interface hypotheses, only the last is capable of explaining the data. Condition and sequence determinants of amyloidogenic propensity that have thus far been reported must be reinterpreted as being reflective of partitioning to hydrophobic-water interfaces. Comparable hydrophobic interfaces are not found in vivo.
authors.affiliation: Department of Chemistry and Chemical Biology, Wright-Rieman Laboratories, Rutgers, the State University of New Jersey, New Brunswick, 610 Taylor Road, Piscataway, New Jersey 08854, USA.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Pronchik
authors.forename: Xianglan
authors.initials: X
authors.lastname: He
authors.forename: Jason T
authors.initials: JT
authors.lastname: Giurleo
authors.forename: David S
authors.initials: DS
authors.lastname: Talaga
mesh_terms: D000595:Amino Acid Sequence; D000682:Amyloid / Q000096:biosynthesis*; D066298:In Vitro Techniques; D018625:Microscopy, Atomic Force; D011108:Polymers / Q000378:metabolism; D051844:alpha-Synuclein / Q000378:metabolism*
chemical_list: D000682:Amyloid; D011108:Polymers; D051844:alpha-Synuclein