Application of confocal single-molecule FRET to intrinsically disordered proteins

Benjamin Schuler, Sonja Müller-Späth, Andrea Soranno, Daniel Nettels

Research output: Chapter in Book/Report/Conference proceedingChapter

25 Scopus citations

Abstract

Intrinsically disordered proteins (IDPs) are characterized by a large degree of conformational heterogeneity. In such cases, classical experimental methods often yield only mean values, averaged over the entire ensemble of molecules. The microscopic distributions of conformations, trajectories, or sequences of events often remain unknown, and with them the underlying molecular mechanisms. Signal averaging can be avoided by observing individual molecules. A particularly versatile method is highly sensitive fluorescence detection. In combination with Förster resonance energy transfer (FRET), distances and conformational dynamics can be investigated in single molecules. This chapter introduces the practical aspects of applying confocal single-molecule FRET experiments to the study of IDPs.

Original languageEnglish
Title of host publicationIntrinsically Disordered Protein Analysis
Subtitle of host publicationVolume 2, Methods and Experimental Tools
EditorsVladimir Uversky, Vladimir Uversky, Keith Dunker
Pages21-45
Number of pages25
DOIs
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
Volume896
ISSN (Print)1064-3745

Keywords

  • Confocal detection
  • Correlation spectroscopy
  • Diffusion
  • FCS
  • FRET
  • Fluorescence spectroscopy
  • Förster resonance energy transfer
  • Intrinsically disordered proteins
  • Photon statistics
  • Single-molecule detection

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  • Cite this

    Schuler, B., Müller-Späth, S., Soranno, A., & Nettels, D. (2012). Application of confocal single-molecule FRET to intrinsically disordered proteins. In V. Uversky, V. Uversky, & K. Dunker (Eds.), Intrinsically Disordered Protein Analysis: Volume 2, Methods and Experimental Tools (pp. 21-45). (Methods in Molecular Biology; Vol. 896). https://doi.org/10.1007/978-1-4614-3704-8_2