Реферат: Incorporation of [2,3,4,5,6-2H5]Phenylalanine, [3,5-2H2]Tyrosine, and [2,4,5,6,7-2H5]Tryptophan into the Bacteriorhodopsin Molecule of Halobacterium halobium

Applied Biochemistry andMicrobiology, Vol. 35, ffo. /. 1999, pp. 29-17. Translated from Prikladnayti Biokhimiya iMikrobialogiya, Vol. 35, No. 1,@  1999, pp. 34-42. Original Russian Text Copyright © /999 hy Mosin, Skluclnev, Shvatz.

Incorporation of [2,3,4,5,6-2H5]Phenylalanine,

[3,5-2H2]Tyrosine,and [2,4,5,6,7-2H5]Tryptophan

into the Bacteriorhodopsin Molecule ofHalobacterium halobium

O. V. Mosin*, D. A. Skladnev**, and V. I. Shvets*

*Lotnonosov Moscow State Academy of Fine Chemical Technology, Moscow, 117571Russia

** StateCenter of Genetics and Selection of Industrial Microorganisms (GNU GENETICA),Moscow, 113515 Russia

Received September 25, 1997

Abstract—Incorporation of[2,3A5,6-2H5]phenylalanine, [3,5-2H2]tyrosine,and [2,4,5,6,7-2H5]tryptophan into thebacteriorhodopsin molecule followed by semipreparative isolation ofbacteriorhodopsin resulted in a yield of 8-10 mg per g bacterial biomass. Thismethod is based on the growth of the strain of halophilic bacteria Halobacteriumhalobium on a synthetic medium containing 2H-labeledaromatic ammo acids and fractionation of solubilized (in 0.5% sodium dodecylsulfate) protein by methanol, including purification of carotenoids. lip-ids, andhigh-molecular-weight and low-molecular-weight compounds, as well asgel-permeation chromatog-raphy on Sephadex G-200. Incorporation of 2H-labeledamino acids was analyzed by electron impact mass spectrometry afterhydrolysis of the protein in 4 N Ba(OH)2 and separation in the formof methyl esters of /V-DNS derivatives of amino aids by re versed-phasehigh-performance liquid chromatography.

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