NMR observation of Tau in Xenopus oocytes.
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.
Bodart
Jean-François
JF
Laboratoire de Régulation des Signaux de division, EA4020, Building SN3, Université des Sciences et Technologies de Lille, 59655 Villeneuve d'Ascq, France.
Wieruszeski
Jean-Michel
JM
Amniai
Laziza
L
Leroy
Arnaud
A
Landrieu
Isabelle
I
Rousseau-Lescuyer
Arlette
A
Vilain
Jean-Pierre
JP
Lippens
Guy
G
D000818:Animals; D005362:Ficoll; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009865:Oocytes / Q000737:chemistry*; D029867:Xenopus Proteins / Q000737:chemistry*; D016875:tau Proteins / Q000737:chemistry*
D009587:Nitrogen Isotopes; D029867:Xenopus Proteins; D016875:tau Proteins; D005362:Ficoll