Eugene Skripkin - Academia.edu (original) (raw)

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Knud Nierhaus

Charité - Universitätsmedizin Berlin / Charité Medical University Berlin

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Papers by Eugene Skripkin

Journal of Biological Chemistry, Dec 1, 1994

Вестник Московского университета. Серия 2: Химия, 1979

Bioorganicheskaya Khimiya, 1981

ABSTRACT

FEBS Letters, Jul 27, 1987

The existence of the internal promoter Pi in the 16 S/23 S intergenic spacers of the rRNA operons... more The existence of the internal promoter Pi in the 16 S/23 S intergenic spacers of the rRNA operons of an eubacterium Escherichia coli and archaebacterium Halobacterium halobium is proposed. The possible functional significance of these promoters is discussed. Internal promoter; rRNA expression; Archaebacteria; rRNA operon; (Halobacterium haIobium, Escherichia co/i)

Journal of the American Chemical Society, Mar 16, 2000

We describe the first direct observation of NsH‚‚‚OdC hydrogen bonding in nucleic acids via the f... more We describe the first direct observation of NsH‚‚‚OdC hydrogen bonding in nucleic acids via the four-bond 4h J NiNj coupling within an N1 i sH1 i ‚‚‚O6 j dC6 j-N1 j segment of a G‚G‚G‚G tetrad. The experiment, two-dimensional (2D) HN(N)-TOCSY, makes use of band-selective 15 N isotropic mixing to transfer magnetization exclusively via the 4h J NN couplings to yield correlations between N1H i (ω 2) and N1 j (ω 1) across the hydrogen-bonded segment. In a complementary experiment, 2D cross-polarization (CP)-H(N)CO-(NN)-TOCSY, employing band-selective heteronuclear 15 N-13 C cross-polarization sequences on either side of the 15 N-15 N TOCSY period, correlations are obtained between N1H i (ω 2) and C6 j (ω 1) nuclei across the hydrogen bond. The symmetric A 2 X 2-type coupling topology of the four-spin nitrogen system in the G‚G‚G‚G tetrad permits accurate measurement of these couplings by a new procedure that fits the experimental data with known analytical isotropic mixing transfer functions. The techniques are demonstrated on a G‚G‚G‚G tetrad formed within a novel dimeric quadruplex fold of a uniformly 13 C/ 15 N-labeled d(G-G-G-T-T-C-A-G-G) DNA sequence. The value of the 4h J NN coupling constant for this system is estimated to be 0.136 (0.021 Hz.

Nucleic Acids Research, 1994

... Alternatively, quantitative data were obtained using a BAS 2000 BlO-Imager (Fuji). ... In fig... more ... Alternatively, quantitative data were obtained using a BAS 2000 BlO-Imager (Fuji). ... In figure 4a, 1 1 n = in ffM ffM When n is plotted as a function of t, a linear plot with a slope of 2 k<iim is expected if the equilibrium model applies. ...

Applied Biochemistry and Biotechnology, Oct 1, 1996

The kinetics of Photinus pyralis and Luciola mingrelica luciferase gene expression was studied on... more The kinetics of Photinus pyralis and Luciola mingrelica luciferase gene expression was studied on plasmids with the thermoinducible XPR promoter in Escherichia coli by SDS-gel electrophoresis of cell lysates to follow luciferase protein-synthesized, enzyme immunoassay (EIA) to follow native enzyme conformer, and the luciferase activity assay. E. coli cells were cultivated at temperature schemes 28-42-21~ or 28-21~ or at alkali pH shift. In the cases of thermoinduction and pH shift, the luciferase expressions have similar features. The 3-h thermoinduction (42~ followed by the incubation at 21~ for 10 h resulted in the maximal amount of the luciferase protein of 4-5% of the total cell proteins. The yield did not change further. The amount of native luciferase conformer and the luciferase activity started to grow after incubation for 10 h at 21~ and reached the maximum after 50-60 h when the synthesized luciferase protein adopted the native-like conformation. At the same time, only 50% of the latter appeared to be catalytically active. An increase in the enzymatic activity correlates with an increase in the intracellular pH and ATP content. Intracellular metabolic reactions were shown to play a role in the conformational changes of the enzyme in a postthermoinduction period, and a possible mechanism of this effect is proposed.

Journal of Biological Chemistry, Dec 1, 1994

Вестник Московского университета. Серия 2: Химия, 1979

Bioorganicheskaya Khimiya, 1981

ABSTRACT

FEBS Letters, Jul 27, 1987

The existence of the internal promoter Pi in the 16 S/23 S intergenic spacers of the rRNA operons... more The existence of the internal promoter Pi in the 16 S/23 S intergenic spacers of the rRNA operons of an eubacterium Escherichia coli and archaebacterium Halobacterium halobium is proposed. The possible functional significance of these promoters is discussed. Internal promoter; rRNA expression; Archaebacteria; rRNA operon; (Halobacterium haIobium, Escherichia co/i)

Journal of the American Chemical Society, Mar 16, 2000

We describe the first direct observation of NsH‚‚‚OdC hydrogen bonding in nucleic acids via the f... more We describe the first direct observation of NsH‚‚‚OdC hydrogen bonding in nucleic acids via the four-bond 4h J NiNj coupling within an N1 i sH1 i ‚‚‚O6 j dC6 j-N1 j segment of a G‚G‚G‚G tetrad. The experiment, two-dimensional (2D) HN(N)-TOCSY, makes use of band-selective 15 N isotropic mixing to transfer magnetization exclusively via the 4h J NN couplings to yield correlations between N1H i (ω 2) and N1 j (ω 1) across the hydrogen-bonded segment. In a complementary experiment, 2D cross-polarization (CP)-H(N)CO-(NN)-TOCSY, employing band-selective heteronuclear 15 N-13 C cross-polarization sequences on either side of the 15 N-15 N TOCSY period, correlations are obtained between N1H i (ω 2) and C6 j (ω 1) nuclei across the hydrogen bond. The symmetric A 2 X 2-type coupling topology of the four-spin nitrogen system in the G‚G‚G‚G tetrad permits accurate measurement of these couplings by a new procedure that fits the experimental data with known analytical isotropic mixing transfer functions. The techniques are demonstrated on a G‚G‚G‚G tetrad formed within a novel dimeric quadruplex fold of a uniformly 13 C/ 15 N-labeled d(G-G-G-T-T-C-A-G-G) DNA sequence. The value of the 4h J NN coupling constant for this system is estimated to be 0.136 (0.021 Hz.

Nucleic Acids Research, 1994

... Alternatively, quantitative data were obtained using a BAS 2000 BlO-Imager (Fuji). ... In fig... more ... Alternatively, quantitative data were obtained using a BAS 2000 BlO-Imager (Fuji). ... In figure 4a, 1 1 n = in ffM ffM When n is plotted as a function of t, a linear plot with a slope of 2 k<iim is expected if the equilibrium model applies. ...

Applied Biochemistry and Biotechnology, Oct 1, 1996

The kinetics of Photinus pyralis and Luciola mingrelica luciferase gene expression was studied on... more The kinetics of Photinus pyralis and Luciola mingrelica luciferase gene expression was studied on plasmids with the thermoinducible XPR promoter in Escherichia coli by SDS-gel electrophoresis of cell lysates to follow luciferase protein-synthesized, enzyme immunoassay (EIA) to follow native enzyme conformer, and the luciferase activity assay. E. coli cells were cultivated at temperature schemes 28-42-21~ or 28-21~ or at alkali pH shift. In the cases of thermoinduction and pH shift, the luciferase expressions have similar features. The 3-h thermoinduction (42~ followed by the incubation at 21~ for 10 h resulted in the maximal amount of the luciferase protein of 4-5% of the total cell proteins. The yield did not change further. The amount of native luciferase conformer and the luciferase activity started to grow after incubation for 10 h at 21~ and reached the maximum after 50-60 h when the synthesized luciferase protein adopted the native-like conformation. At the same time, only 50% of the latter appeared to be catalytically active. An increase in the enzymatic activity correlates with an increase in the intracellular pH and ATP content. Intracellular metabolic reactions were shown to play a role in the conformational changes of the enzyme in a postthermoinduction period, and a possible mechanism of this effect is proposed.

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