NMR observation of Tau in Xenopus oocytes

J Magn Reson. 2008 Jun;192(2):252-7. doi: 10.1016/j.jmr.2008.03.006. Epub 2008 Mar 14.

Abstract

The observation by NMR spectroscopy of microinjected 15N-labelled proteins into Xenopus laevis oocytes might open the way to link structural and cellular biology. We show here that embedding the oocytes into a 20% Ficoll solution maintains their structural integrity over extended periods of time, allowing for the detection of nearly physiological protein concentrations. We use these novel conditions to study the neuronal Tau protein inside the oocytes. Spectral reproducibility and careful comparison of the spectra of Tau before and after cell homogenization is presented. When injecting Tau protein into immature oocytes, we show that both its microtubule association and different phosphorylation events can be detected.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Ficoll
  • Nitrogen Isotopes
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Oocytes / chemistry*
  • Xenopus Proteins / chemistry*
  • tau Proteins / chemistry*

Substances

  • Nitrogen Isotopes
  • Xenopus Proteins
  • tau Proteins
  • Ficoll