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81.
In their inhibition-inducing interactions with enzymes, quinones primarily utilize two mechanisms, arylation and oxidation of enzyme thiol groups. In this work, we investigated the interactions of 1,4-naphthoquinone with urease in an effort to estimate the contribution of the two mechanisms in the enzyme inhibition. Jack bean urease, a homohexamer, contains 15 thiols per enzyme subunit, six accessible under non-denaturing conditions, of which Cys592 proximal to the active site indirectly participates in the enzyme catalysis. Unlike by 1,4-benzoquinone, a thiol arylator, the inactivation of urease by 1,4-naphthoquinone under aerobic conditions was found to be biphasic, time- and concentration-dependent with a non-linear residual activity-modified thiols dependence. DTT protection studies and thiol titration with DTNB suggest that thiols are the sites of enzyme interactions with the quinone. The inactivated enzyme had approximately 40% of its activity restored by excess DTT supporting the presence of sulfenic acid resulting from the oxidation of enzyme thiols by ROS. Furthermore, the aerobic inactivation was prevented in approximately 30% by catalase, proving the involvement of hydrogen peroxide in the process. When H2O2 was directly applied to urease, the enzyme showed susceptibility to this inactivation in a time- and concentration-dependent manner with the inhibition constant of H2O2 Ki = 3.24 mM. Additionally, anaerobic inactivation of urease was performed and was found to be weaker than aerobic. The results obtained are consistent with a double mode of 1,4-naphthoquinone inhibitory action on urease, namely through the arylation of the enzyme thiol groups and ROS generation, notably H2O2, resulting in the oxidation of the groups. 相似文献
82.
Blanka Orłowska Ewa Augustynowicz-Kopeć Monika Krajewska Anna Zabost Mirosław Welz Stanisław Kaczor Krzysztof Anusz 《European Journal of Wildlife Research》2017,63(1):21
The aim of this study was to assess whether animal tuberculosis (TB) is transmitted between free-living European bison (Bison bonasus caucasicus), wild boars (Sus scrofa), and protected carnivores such as grey wolves (Canis lupus), brown bears (Ursus arctos), and Eurasian lynx (Lynx lynx) in the Bieszczady Mountains in Southern Poland. Results of animal studies suggest that TB transmission from bison or wild boars to grey wolves is possible. These are the first described cases where Mycobacterium caprae was detected in samples collected from grey wolves. 相似文献
83.
W M Krajewska A Lipińska M Gaczyński L K?yszejko-Stefanowicz 《The International journal of biochemistry》1992,24(5):759-767
- 1.1. As a further step toward characterizing nonhistone protein of mol. wt 48,000 which was found to be much more abundant in animal tumour cells than in normal ones [Krajewska W.M., Lipinska A., Marszatek M., Kiliańska Z., Wojtkowiak Z. and Kłyszejko-Stefanowicz L. Cell. Biochem. Funct. 8, 79–89 (1990)] its intranuclear localization in hamster liver and Kirkman-Robbins hepatoma was studied. The protein was identified by immunoblotting technique in the presence of antibodies against polypeptide of mol. wt about 48,000 from Kirkman-Robbins hepatoma.
- 2.2. Distribution of antigen with mol. wt of 48,000 in nuclear fractions representing different levels of nuclear material organization i.e. in nucleoli, nuclease-sensitive and nuclease-resistant fractions, and extensive nuclease digestion products separated by size on Bio-Gel A-50m, implied the structural role of this component.
- 3.3. Fractionation of endogenously digested nuclei into low salt extract, high salt extract and nuclear matrix revealed that in normal liver the antigen studied is associated with nuclear matrix while in hepatoma this component appeared in high salt extract.
- 4.4. These results suggest that polypeptide with mol. wt of 48,000 is a shuttling protein which may be involved in reorganization of nuclear matrix during neoplastic transformation.
84.
Z Kilianska W M Krajewska M Gaczynski M Marszalek L Klyszejko-Stefanowicz 《The International journal of biochemistry》1989,21(8):873-881
1. Non-histone chromatin protein fractions NHCP1 and NHCP2 eluted from hydroxyapatite with 50 and 100 mM phosphate buffer (pH 6.8) from nuclei of Kirkman-Robbins hepatoma from 4th, 7th and 9th day of growth were analysed by one- and two-dimensional gel electrophoresis as well as Western blot technique in the presence of antibodies elicited against NHCP1, NHCP2 and dehistonized chromatin of hamster hepatoma and liver. 2. The presence of electrophoretically and immunologically specific components among NHCP1 and NHCP2 fractions during Kirkman-Robbins hepatoma growth was stated. 相似文献
85.
Wies
awa Zaborska Barbara Krajewska Maciej Leszko Zofia Olech 《Journal of Molecular Catalysis .B, Enzymatic》2001,13(4-6):103-108
The inhibition of jack bean urease by Ni2+ ions was studied in 20 mM HEPES buffer pH 7.0. The inhibition was observed in two systems which differed in the order in which the components of the reaction mixture were mixed. In the first (unincubated), the reaction was initiated by adding urease to the mixture of urea and Ni2+ ions, and in the second (incubated), by adding urea to the mixture of urease incubated with Ni2+ ions prior to the reaction. It was shown that Ni2+ ions are a competitive slow-binding inhibitor of urease. In the first system the inhibition constants are Ki=0.042 mM and Ki*=0.0028 mM, and in the second system Ki*=0.0024 mM. The inhibition was found to involve the rapid formation of a urease-Ni2+complex followed by its relatively slow, reversible isomerization, with forward and reverse rate constants of 0.64 and 0.045 min−1, respectively. 相似文献
86.
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88.
W M Krajewska L K?yszejko-Stefanowicz 《Comparative biochemistry and physiology. B, Comparative biochemistry》1982,71(1):145-148
1. Chromatin proteins of chicken thrombocytes and erythrocytes were separated into three fractions by successive extraction with 5 M urea containing various salt concentrations and pH values. Molecular composition of protein fractions was determined by SDS-polyacrylamide gel electrophoresis. 2. The efficiences of the chromatin residues after sequential protein extractions as well as those of reconstituted DNA-protein fraction complexes, in serving as a template for the in vitro RNA synthesis were measured in order to identify the effect of each fraction. 3. The different involvement of chromatin protein fractions of template properties of thrombocyte and erythrocyte chromatin was stated. 相似文献
89.
90.