Functional Expression and Properties of Sec14p-Like Protein with Molecular Mass 45 kD from Rat Olfactory Epithelium

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V. V. Radchenko^1*, M. I. Merkulova¹, T. M. Shuvaeva¹, T. N. Simonova¹, A. A. Bondar², and V. M. Lipkin¹

¹Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997 Moscow, Russia; fax: (7-095) 335-7103; E-mail: radchenko@mail.ibch.ru

²Institute of Cell Biophysics, Russian Academy of Sciences, 142292 Pushchino, Moscow Region, Russia; fax: (7-096) 779-0509; E-mail: novoselov@mail.icb.psn.ru

^* To whom correspondence should be addressed.

Received March 16, 2005; Revision received June 1, 2005

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cDNA of Sec14p-like water-soluble protein with molecular mass 45 kD from rat olfactory epithelium was expressed in Escherichia coli Rosetta^TM cells. The expression product was purified by a two-step chromatographic procedure on DEAE-Sepharose and Sephacryl S-200. The identity of structural and functional characteristics of the recombinant and native proteins was demonstrated by CD, mass spectrometry, and Western blotting. Using several lipids immobilized on nitrocellulose membranes, it was shown that phosphatidylinositol-3,4,5-triphosphate is the specific ligand for the studied protein.

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KEY WORDS: lipid-binding protein p45, olfactory epithelium, Sec14p family, CRAL/TRIO domain, protein-lipid interaction, phosphatidylinositol-3,4,5-triphosphate

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