- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
FerB is a flavoenzyme capable of reducing quinones, ferric complexes and chromate. Its expression in Escherichia coli as a hexahistidine fusion resulted in a functional product only when the tag was placed on the C-terminus. The molecular mass values estimated by gel permeation chromatography were compatible with the existence of either dimer or trimer, whereas the light scattering data, together with cross-linking experiments that yielded exclusively monomer and dimer bands on dodecyl sulfate-polyacrylamide gels, strongly supported a dimeric nature of both native and tagged form of FerB. These two proteins also exhibited almost identical secondary structure as judged by Fourier transform infra red spectrometry. The presence of tag, however, shifted the temperature of thermal inactivation as well as the thermal denaturation curve towards lower temperatures. Despite somewhat lower thermal stability, the fusion protein is considered a better candidate for crystallization than the wild-type one due to a more negative value of its second optical viral coefficient.
- MeSH
- diferenciální skenovací kalorimetrie MeSH
- Escherichia coli genetika MeSH
- Fourierova analýza MeSH
- histidin genetika chemie metabolismus MeSH
- multimerizace proteinu MeSH
- NADH, NADPH oxidoreduktasy biosyntéza genetika chemie metabolismus MeSH
- NADP metabolismus MeSH
- oligopeptidy genetika chemie metabolismus MeSH
- Paracoccus denitrificans enzymologie genetika MeSH
- rekombinantní fúzní proteiny biosyntéza genetika chemie metabolismus MeSH
- sekundární struktura proteinů MeSH
- stabilita enzymů MeSH
- teplota MeSH
- Publikační typ
- práce podpořená grantem MeSH
A new hydrophobic platinum(IV) complex, LA-12, a very efficient anticancer drug lacking cross-resistance with cisplatin (CDDP), is now being tested in clinical trials. Here we investigated the apoptogenic activity of LA-12 and its effect on gap-junctional intercellular communication (GJIC) in the rat liver epithelial cell line WB-F344. LA-12 induced apoptosis much more efficiently than did CDDP due to a combination of rapid penetration into the cell and attack on DNA, leading to fast activation of p53 and caspase-3. Exposure of WB-F344 cells to LA-12 led to rapid induction of the time- and dose-dependent decrease in GJIC. On the molecular level, loss of GJIC induced by LA-12 was mediated by activation of extracellular signal-regulated kinase (ERK)-1 and ERK-2, as demonstrated by the use of inhibitors of ERK activation. Inhibition of GJIC was linked to rapid hyperphosphorylation of connexin-43 and disappearance of connexon clusters from membranes, which was not observed in the case of CDDP.
- MeSH
- amantadin analogy a deriváty farmakologie MeSH
- antitumorózní látky farmakologie MeSH
- apoptóza MeSH
- buněčné linie MeSH
- cisplatina farmakologie MeSH
- epitelové buňky metabolismus MeSH
- financování organizované MeSH
- fosforylace MeSH
- hydrofobní a hydrofilní interakce MeSH
- konexin 43 metabolismus MeSH
- krysa rodu rattus MeSH
- mezerový spoj účinky léků MeSH
- nádorové buněčné linie MeSH
- organoplatinové sloučeniny farmakologie MeSH
- sloučeniny platiny chemie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- nemoci prasat imunologie MeSH
- nukleokapsida genetika MeSH
- otevřené čtecí rámce genetika imunologie MeSH
- specificita protilátek MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- kongresy MeSH
