Thioflavin-S staining coupled to flow cytometry. A screening tool to detect in vivo protein aggregation

Mol Biosyst. 2012 Nov;8(11):2839-44. doi: 10.1039/c2mb25214g. Epub 2012 Aug 7.

Abstract

Amyloid deposits are associated with an increasing number of human disorders, including Alzheimer's and Parkinson's diseases. Recent studies provide compelling evidence for the existence of amyloid-like conformations in the insoluble bacterial inclusion bodies (IBs) produced during the recombinant expression of amyloidogenic proteins. This makes prokaryotic cells a physiologically relevant system to study the mechanisms of in vivo amyloid deposition. We show here that the application of flow cytometry to detect Thioflavin-S (Th-S) fluorescence provides a fast, robust, quantitative, non-invasive method to screen for the presence of in vivo intracellular amyloid-like aggregates in bacteria, with potential application in the analysis of the impact of genetic mutations or chemical compounds on the aggregation of disease-associated polypeptides.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amyloid / metabolism
  • Benzothiazoles
  • Flow Cytometry / methods*
  • Inclusion Bodies / metabolism
  • Protein Binding
  • Proteins / metabolism*
  • Thiazoles / chemistry*

Substances

  • Amyloid
  • Benzothiazoles
  • Proteins
  • Thiazoles
  • thioflavin T