Functional expression and properties of Sec14p-like protein with molecular mass 45 kD from rat olfactory epithelium

cDNA of Sec14p-like water-soluble protein with molecular mass 45 kD from rat olfactory epithelium was expressed in Escherichia coli Rosetta cells. The expression product was purified by a two-step chromatographic procedure on DEAE-Sepharose and Sephacryl S-200. The identity of structural and functio...

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Bibliographic Details
Published in:Biochemistry (Moscow) 2005-12, Vol.70 (12), p.1341-1347
Main Authors: Radchenko, V V, Merkulova, M I, Shuvaeva, T M, Simonova, T N, Bondar, A A, Lipkin, V M
Format: Article
Language:English
Subjects:
Animals
Blotting, Western
Carrier Proteins - biosynthesis
Carrier Proteins - isolation & purification
Carrier Proteins - metabolism
Circular Dichroism
Cloning, Molecular
E coli
Escherichia coli
Escherichia coli - metabolism
Ligands
Lipids
Mass Spectrometry
Molecular Weight
Olfactory Mucosa - metabolism
Olfactory receptors
Phosphates
Phosphatidylinositol
Phosphatidylinositol Phosphates - metabolism
Proteins
Rats
Rats, Wistar
Recombinant Proteins - biosynthesis
Recombinant Proteins - isolation & purification
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Summary:cDNA of Sec14p-like water-soluble protein with molecular mass 45 kD from rat olfactory epithelium was expressed in Escherichia coli Rosetta 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.
ISSN:0006-2979
1608-3040
0320-9725
DOI:10.1007/s10541-005-0267-0