DETERMINATION OF STRUCTURE AND FUNCTIONAL PROPERTIES OF CYTOPLASMIC TERMINUS OF VANILLOID RECEPTOR TRPV1

Veronika Mrazikova¹, Eva Jindrova¹, Viktorie Vlachova², Rudiger Ettrich¹ and Jan Teisinger²

¹Laboratory of High Performance Computing, Institute of Physical Biology USB and

Institute of Landscape Ecology AS CR, Zamek 136, CZ-373 33 Nove Hrady, Czech Republic

²Institute of Physiology, Academy of Sciences, 142 20 Prague 4, Czech Republic

The vanilloid receptor TRPV1, a member of TRP channel family, has function as a multimodal signal transducter of noxious stimuli in the mammalian somatosensory system [1]. The TRPV1 is consisted of six transmembarane-spanning domains with a pore forming region between fifth and sixth domains, and cytoplasmically located C- and N-terminal regions. Although structural and functional studies have been done [2, 3], the possible contributions of terminal regions to vanilloid receptor function remain elusive. To determine structure and functional properties of the cytoplasmically located tails, the DNA fragments encoding for the N- and C- terminus were cloned to the expression vectors and transformed to E. coli strain. Overexpressed proteins were purified by affinity chromatography and used for structural analysis by a wide range of low resolution methods. Experimental results were combined with homology and energetic modeling techniques and we propose a three-dimensional structure of the C-terminus.

1. M.J. Caterina, M.A. Schumacher, M. Tominaga, T.A. Rosen, J.D. Levine, D. Julius, Nature, 389 (1997) 816-824.

2. V. Vlachová, J. Teisinger, K. Susánková, A. Lyfenko, R. Ettrich, and L. Vyklický, J. Neurosci., 23(4) (2003) 1340-1350.

3. N. García-Sanz, A. Fernández-Carvajal, C. Morenilla-Palao, R. Planells-Cases, E. Fajardo-Sánchez, G. Fernández-Ballester, and A. Ferrer-Montiel, J. Neurosci., 24(23) (2004) 5307-5314.

This research was supported by the Ministry of Education, Youth and Sports of the Czech Republic (MSM6007665808) and by the Academy of Sciences of the Czech Republic (Institutional research concepts AVOZ50110509 and AVOZ60870520).