Simultaneous measurement of amyloid fibril formation by dynamic light scattering and fluorescence reveals complex aggregation kinetics

PLoS One. 2013;8(1):e54541. doi: 10.1371/journal.pone.0054541. Epub 2013 Jan 17.

Abstract

An apparatus that combines dynamic light scattering and Thioflavin T fluorescence detection is used to simultaneously probe fibril formation in polyglutamine peptides, the aggregating subunit associated with Huntington's disease, in vitro. Huntington's disease is a neurodegenerative disorder in a class of human pathologies that includes Alzheimer's and Parkinson's disease. These pathologies are all related by the propensity of their associated protein or polypeptide to form insoluble, β-sheet rich, amyloid fibrils. Despite the wide range of amino acid sequence in the aggregation prone polypeptides associated with these diseases, the resulting amyloids display strikingly similar physical structure, an observation which suggests a physical basis for amyloid fibril formation. Thioflavin T fluorescence reports β-sheet fibril content while dynamic light scattering measures particle size distributions. The combined techniques allow elucidation of complex aggregation kinetics and are used to reveal multiple stages of amyloid fibril formation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alzheimer Disease* / metabolism
  • Alzheimer Disease* / pathology
  • Amino Acid Sequence
  • Amyloid / biosynthesis
  • Amyloid / ultrastructure*
  • Amyloid beta-Peptides / chemistry
  • Amyloid beta-Peptides / metabolism
  • Benzothiazoles
  • Fluorescence
  • Humans
  • Huntington Disease* / metabolism
  • Huntington Disease* / pathology
  • Kinetics
  • Light
  • Parkinson Disease* / metabolism
  • Parkinson Disease* / pathology
  • Particle Size
  • Protein Structure, Secondary
  • Thiazoles / chemistry

Substances

  • Amyloid
  • Amyloid beta-Peptides
  • Benzothiazoles
  • Thiazoles
  • thioflavin T