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15N magnetic relaxation study of backbone dynamics of the ribosome-associated cold shock response protein Yfia of Escherichia coli

100%
Zhukov I., Bayer P., Schölermann B., Ejchart A.
Acta Biochimica Polonica
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2007
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vol. 54
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issue 4
769-775
EN
In the solution structure of the ribosome-associated cold shock response protein Yfia of Escherichia coli in the free state two structural segments can be distinguished: a well structured, rigid N-terminal part displaying a βαβββα topology and a flexible C-terminal tail comprising last 20 amino-acid residues. The backbone dynamics of Yfia protein was studied by 15N nuclear magnetic relaxation at three magnetic fields and analyzed using model-free approach. The overall diffusional tumbling of the N-terminal part is strongly anisotropic with a number of short stretches showing increased mobility either on a subnanosecond time scale, or a micro- to millisecond time scale, or both. In contrast, the unstructured polypeptide chain of the C-terminal part, which cannot be regarded as a rigid structure, shows the predominance of fast local motions over slower ones, both becoming faster closer to the C-terminus.
2
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Apoptosis induced by membrane damage in human lymphocytes; effects of arachidonic acid and its photoproducts.

100%
Zarębska Z., Zielińska J., Zhukov I., Maśliński W.
Acta Biochimica Polonica
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2005
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vol. 52
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issue 1
179-194
EN
The effect of arachidonic acid (AA) combined with UVA irradiation was studied in a model system mimicking phototherapy PUVA (psoralen+UVA) ex vivo in vitro. The contribution of damage to the plasma membrane by PUVA was tested on human lymphocytes derived from healthy donors. The effect of arachidonic acid (AA) combined with UVA irradiation was compared with that of a psoralen photoadduct to AA added to the culture. The adduct, obtained photochemically and purified, was characterized by NMR and MS spectrometry as a cycloadduct of psoralen to the vinylene bond of the acid (AA<>PSO). The reactions of cultured cells, manifested 20 h after treatment by changes in apoptosis and mitochondrial depolarization, were monitored by flow cytometry by tagging lymphocytes with appropriate fluorescent probes. Treatment of lymphocyte suspension within AA doses from 40 to 100 μM gradually induced a shift from Anx-V+ (single positive cells) to late apoptotic, Anx-V+PI+ (double positive cells) in a dose dependent manner. The adduct, AA<>PSO, induced apoptotic changes at a concentration 2-3 times higher than free AA. Combination of psoralen (1 μM ) or arachidonic acid (20-120 μM) with UVA irradiation (2-6 J/cm2) accelerated the plasma membrane changes in a synergic way. Preliminary studies indicated that changes in the transmembrane potential of mitochondria paralleled the apoptosis when cells were treated by AA alone. Our findings showed that UVA radiation of lymphocytes in the presence of arachidonic acid, as in the presence of psoralen, enhanced apoptosis of cells in a synergic manner. Thus, PUVA-induced apoptosis may proceed in part by a still undefined signaling pathway(s) triggered in lymphocyte membranes.
3
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NMR measurements of proton exchange between solvent and peptides and proteins

88%
Wójcik J., Ruszczyńska K., Zhukov I., Ejchart A.
Acta Biochimica Polonica
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1999
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vol. 46
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issue 3
651-663
4
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Factors improving the accuracy of determination of 15N relaxation parameters in proteins

87%
Zhukov I., Ejchart A.
Acta Biochimica Polonica
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1999
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vol. 46
|
issue 3
665-671
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