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In vitro polymerization of tau protein monitored by laser light scattering: method and application to the study of FTDP-17 mutants.

Abstract
Tau polymerization into the filaments that compose neurofibrillary tangles is seminal to the development of many neurodegenerative diseases. It is therefore important to understand the mechanisms involved in this process. However, a consensus method for monitoring tau polymerization in vitro has been lacking. Here we demonstrate that illuminating tau polymerization reactions with laser light and measuring the increased scattering at 90 degrees to the incident beam with a digital camera results in data that closely approximate the mass of tau polymer formation in vitro. The validity of the technique was demonstrated over a range of tau concentrations and through multiple angle scattering measurements. In addition, laser light scattering data closely correlated with quantitative electron microscopy measurements of the mass of tau filaments. Laser light scattering was then used to measure the efficiency with which the mutant tau proteins found in frontotemporal dementia and Parkinsonism linked to chromosome 17 (FTDP-17) form filamentous structures. Several of these mutant proteins display enhanced polymerization in the presence of arachidonic acid, suggesting a direct role for these mutations in tau the filament formation that characterizes FTDP-17.
AuthorsT C Gamblin, M E King, H Dawson, M P Vitek, J Kuret, R W Berry, L I Binder
JournalBiochemistry (Biochemistry) Vol. 39 Issue 20 Pg. 6136-44 (May 23 2000) ISSN: 0006-2960 [Print] United States
PMID10821687 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)
Chemical References
  • tau Proteins
Topics
  • Chromosomes, Human, Pair 17
  • Dementia (genetics)
  • Frontal Lobe (chemistry)
  • Humans
  • Lasers
  • Light
  • Microscopy, Electron
  • Mutagenesis, Site-Directed
  • Mutation, Missense
  • Parkinsonian Disorders (genetics)
  • Reproducibility of Results
  • Scattering, Radiation
  • Sensitivity and Specificity
  • Temporal Lobe (chemistry)
  • tau Proteins (chemistry, genetics, metabolism, ultrastructure)

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