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1

Factors improving the accuracy of determination of 15N relaxation parameters in proteins

100%
Zhukov I., Ejchart A.
Acta Biochimica Polonica
|
1999
|
tom 46
|
nr 3
A number of factors at all stages of data processing which affect the accuracy of determination of 15N relaxation parameters in 15N-labeled proteins is discussed. Methods which allow to improve accuracy of the determined parameters are presented using data obtained for Cucurbita maxima trypsin inhibitor.
2

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

81%
Zhukov I., Bayer P., Scholermann B., Ejchart A.
Acta Biochimica Polonica
|
2007
|
tom 54
|
nr 4
769-775
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 over all 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 milli second 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.
3

PUVA-induced lipoapoptosis in human lymphocytes: arachidonic acid and its photoproducts

81%
Zarebska Z., Zielinska J., Zhukov I., Maslinski W.
Acta Biochimica Polonica
|
2003
|
tom 50
|
nr Supplement 1
4

Apoptosis induced by membrane damage in human lymphocytes; effects of arachidonic acid and its photoproducts

81%
Zarebska Z., Zielinska J., Zhukov I., Maslinski W.
Acta Biochimica Polonica
|
2005
|
tom 52
|
nr 1
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.
5

NMR measurements of proton exchange between solvent and peptides and proteins

81%
Wojcik J., Ruszczynska K., Zhukov I., Ejchart A.
Acta Biochimica Polonica
|
1999
|
tom 46
|
nr 3
Scope and limitations of the NMR based methods, equilibration and magnetization transfer, for measuring proton exchange rates of amide protons in peptides and proteins with water protons are discussed. Equilibration is applied to very slow processes detected by hydrogen-deuterium exchange after a solute is dissolved in D2O. Magnetization transfer allows to study moderately rapid processes in H2O. A number of precautions should be undertaken in order to avoid systemic errors inherent in the magnetization transfer method.
6

Expression and structural studies on hepatitis C virus helicase and its inhibitors

51%
Stankiewicz A., Strokovskaya L., Poznanski J., Zhukov I., Gozdek A., Bretner M., Zagorska-Ostoja W., Boguszewska-Chachulska A.
Acta Biochimica Polonica
|
2003
|
tom 50
|
nr Supplement 1
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