Статья: Получение мембранного белка бактериородопсина, меченного дейтерием по остаткам ароматических аминокислот L-фенилаланина, L-тирозина и L-триптофана
1. Oesterhelt D. and Stoeckenius W. // Nature New Biol. - 1971. - V.233. - P.149.
2. Spudich John L. // Ann. Rev. Biophys. Chem. - 1988. - V.17. - P.193-215.
3. Creuzet F., McDermott A., Gebhard R., Hoef K., van der Spijker, Assink M. M., Herzfeld J., Lugtenburg J., Levitt M.H., Griffin R.G. // Science. - 1991. - V.251. - P.250-286.
4. Argade P.V., Kenneth J., Rothscild K.J., Kawamoto A.H., Herzfeld J., and Herlihy W.C. // Proc. Natl. Acad. Sci. USA. -1981. - V.78. - No.3. - P.1643-1646.
5. Rothschild K.J., Braiman M.S., Yi-Wu He, Marti T. and Khorana H. G. // J. of Biological Chemistry. - 1990. - P.16985-16990.
6. Otto H., Marti T., Holz M., Mogi T., Stern L.J., Engel F., Khorana H.G. & Heyn M.P. // Proc. Natl. Acad. Sci. U.S.A. - 1990. - V.87. - P.1018-1022..
7.Vetter W. , in: Biochemical Applications of mass-spectrometry (Walles G.R., and Dormor O.C.). - 1980. -First supplementary volume. - Wiley - Interscience. - N.Y. - USA. - P.439.
8. Hackett N.R., Stern L.J., Chao B.H., Kronis K.A. and Khorana H.J. // J. Biol. Chem. - 1987. - V.262. - P.9270-9277.
9. Rosenbach V., Goldberg R., Gilon C. and Ottolenghi M. // Photochem. Photobiol. - 1982. - V.36. - P.197-201.
10. Plotkin B.J., Sherman M.V. // Biochem. - 1984. - V.23. - P.5353-5360.
11. Griffiths D.V., Feeney J., Roberts G.C.and Burgen A.S. // Biochim. et Biophys. Acta. - 1976. - V.446. - P.479-585.
12.Matthews H.R., Kathleen S., Matthews K. and Stanley J. // Biochim. et Biophys. Acta. - 1977. - V.497. P.1-13.
13. Onishi H., McCance M.E., and Gibbons N.E. // Canad. J. of Microbiol. - 1965. - V.11. - P.365-373.
14. Oesterhelt D., Hess B. // Eur. J. Biochem. - 1973. - V.37. - N.1. - P.316-326.
15. Tokunada F., Ebrey T. . // Biochemistry. - 1978. - V.17. - N.10. - P.1915-1922..
16. Гринштейн Дж, Виниц М. Химия аминокислот и пептидов. - М.:Мир. - 1965. - С. 387-390. 16.
17. Егорова Т.А., Мосин О.В., Еремин С.В., Карнаухова Е.Н., Звонкова Е.Н., Швец В.И. // Биотехнология. - 1993. - Т.8. - С.21-24.
18. Oesterhelt D. & Stoeckenius W. // Methods Enzymol. - 1974. - V.31. - P.667-678.
19. Звонкова Е.Н., Зотчик Н.В., Филлипович Е.И., Митрофанова Т.К., Мягкова Г.И., Серебренникова Г.А // Химия биологически активных природных соединений. - М.: Химия, 1970. - С.65-68.
20. Cohen J.S., Putter I. // Biochim. Biophys. Acta. - 1970. - V.222. - P.515-520.
O.V. МОSIN
Moscow State Academy of Fine Chemical Technology named after M.V. Lomonosov, 117571.
PREPARATION OF BACTERIORHODOPSIN LABELED WITH DEUTERIUM ON AROMATIC RESIDUES OF AROMATIC AMINO ACIDS L-PHENYLALANINE, L-TYROSINE AND L-TRYPTOPHAN.
The samples of membrane protein bacteriorhodopsin wich contained selective deuterated analoques of aromatic amino acids such as L-phenylalanine, L-tyrosine and L-triptophan were obtained biosynthetically. It was achieved through the cultivation of a strain of halophilic bacterium H. halobium ЕТ 1001 on synthetic medium containing instead of natural amino acids the chemically synthetized L-[2,3,4,5,6-2 H5 ]-phenylalanine, L-[3,5-2 H2 ]-tyrosine and L-[2,4,5,6,7-2 H5 ]-tryptophan. The data on cultivation of H. halobium on media containing the deuterated analoques of amino acids and preparation of deuterared bacteriorhodopsin are presented. The analysis of deuterium enrichment of amino acids obtained from hydrolizates of bacteriorhodopsin was perfomed by the electron impact mass-spectrometry method of methyl esters of dansyl-amino acids and benzyloxycarbonyl- amino acid derivatives after preparative separation using reverse-phase high perfomance liquid chromatofraphy (HPLC). It was shown, that in mass spectra of derivatives of amino acids obtained from hydrolizates of bacteriorhodopsin molecular ions of deuterated aromatic amino acids are presented and practically absent their native non-labelled analoques. The data obtained testified to the high efficiency of labelling of bacteriorhodopsin in those experimental conditions.