In-cell NMR in Leishmania tarentolae

S. Trantirkova¹, V. Dotsch², K. Alexandrov³, R. Breitling³, L. Trantirek¹

¹University of South Bohemia, Faculty of Biological Sciences, Ceske Budejovice (Czech Republic)

² W. Goethe University, Institute for Biophysical Chemistry and Center for Biomolecular Magnetic Resonance, Frankfurt (Germany)

³Planck Institute for Molecular Physiology, Department of Physical Biochemistry, Dortmund (Germany)

trant@bf.jcu.cz

To assess the influence of the native-like environment and a drug binding on a protein tertiary structure, we are developing a methodology for aquiring in vivo high-resolution NMR spectra of the proteins over-expressed inside L. tarentolae cells. L. tarentolae, in contrast to other expression systems such as E. coli or baculovirus, possess manmalian-type of post-translational modifications and full protein folding machinery. The parameters critical for observing proteins inside living L. tarentolae cells using NMR spectroscopy are evaluated.

This work was supported by Ministry of Education of the Czech Republic (1K04011, MSM6007665801) and by Grant Agency of the Czech Republic (204/04/P191).