Journal
JOURNAL OF MAGNETIC RESONANCE
Volume 192, Issue 2, Pages 252-257Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jmr.2008.03.006
Keywords
NMR; in-cell; Xenopus oocytes; ficoli; cell survival; Tau; phosphorylation
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The observation by NMR spectroscopy of microinjected N-15-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. (c) 2008 Elsevier Inc. All rights reserved.
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