Differential protein synthesis after red light illuminations in germinating fern spores

The effects of red and far red illuminations were studied in spores of the dark germinatingPteridium aquilinum and that of the light dependently germinatingDryopteris filix-mas. Proteins were analysed by electrophoresis according to their molecular weights. The isoesterases were separated by gel electrofocusing. In both species we found two newly synthesized protein bands, probably controlled by the presence of active phytochrome. One of these seems to be identical, the other is different in the two species. Phytochrome control was also detected in the esterase isoenzyme pattern.     

Urethane and hydroxyurethane induce sister-chromatid exchanges in cultured human lymphocytes.

Both urethane and hydroxyurethane induced sister-chromatid exchanges (SCE) in cultured human lymphocytes. Aroclor-induced rat-liver microsome fraction deactivated rather than activated these two agents in the lymphocyte system.     

Temporal variation in juvenile fish communities of Kaunas reservoir littoral zone,Lithuania

The aim of this study was to assess juvenile fish communities in terms of species composition, fish diversity and density in the littoral zone of the Kaunas reservoir before (in 1989–1990, period I) and after (in 1999–2000, period II, and in 2006–2007, period III) launching the Kruonis hydroelectric pumped plant (Kruonis HPP). During the whole research period, 20 fish species were caught. According to the frequency of occurrence, the three-spined stickleback Gasterosteus aculeatus, European perch Perca fluviatilis and roach Rutilus rutilus were regarded as constant species in all investigated periods. Significant differences were established in juvenile fish community density between period I and periods II and III, whereas species richness (S) and species diversity indices (H′, J′) did not change significantly. The density of the shoreline community in period III was more than two times lower than in period I, probably due to higher fluctuations in water level of the reservoir, resulting from the Kruonis HPP operation.     

Probing the conformational changes and peroxidase-like activity of cytochrome c upon interaction with iron nanoparticles

Herein, the interaction of iron nanoparticle (Fe-NP) with cytochrome c (Cyt c) was investigated, and a range of techniques such as dynamic light scattering (DLS), zeta potential measurements, static and synchronous fluorescence spectroscopy, near and far circular dichroism (CD) spectroscopy, and ultraviolet–visible (UV–vis) spectroscopy were used to analyze the interaction between Cyt c and Fe-NP. DLS and zeta potential measurements showed that the values of hydrodynamic radius and charge distribution of Fe-NP are 83.95 ± 3.7 nm and 4.5 ± .8 mV, respectively. The fluorescence spectroscopy results demonstrated that the binding of Fe-NP with Cyt c is mediated by hydrogen bonds and van der Waals interactions. Also Fe-NP induced conformational changes in Cyt c and reduced the melting temperature value of Cyt c from 79.18 to 71.33°C. CD experiments of interaction between Fe-NP and Cyt c revealed that the secondary structure of Cyt c with the dominant α-helix structures remained unchanged whereas the tertiary structure and heme position of Cyt c are subjected to remarkable changes. Absorption spectroscopy at 695 nm revealed that Fe-NP considerably disrupt the Fe…S(Met80) bond. In addition, the UV–vis experiment showed the peroxidase-like activity of Cyt c upon interaction with Fe-NP. Hence, the data indicate the Fe-NP results in unfolding of Cyt c and subsequent peroxidase-like activity of denatured species. It was concluded that a comprehensive study of the interaction of Fe-NP with biological system is a crucial step for their potential application as intracellular delivery carriers and medicinal agents.     

Bacterial fermentation and isotope labelling optimized for amyloidogenic proteins

We developed a cost sensitive isotope labelling procedure using a fed-batch fermentation method and tested its efficiency producing the ¹⁵N-, ¹³C- and ¹⁵N/¹³C-labelled variants of an amyloidogenic miniprotein (E5: EEEAVRLYIQWLKEGGPSSGRPPPS). E5 is a surface active protein, which forms amyloids in solution. Here, we confirm, using both PM-IRRAS and AFM measurements, that the air–water interface triggers structural rearrangement and promotes the amyloid formation of E5, and thus it is a suitable test protein to work out efficient isotope labelling schemes even for such difficult sequences. E. coli cells expressing the recombinant, ubiquitin-fused miniprotein were grown in minimal media containing either unlabelled nutrients, or ¹⁵N-NH₄Cl and/or ¹³C-D-Glc. The consumption rates of NH₄Cl and D-Glc were quantitatively monitored during fermentation and their ratio was established to be 1:5 (for NH₄Cl: D-Glc). One- and two-step feeding schemes were custom-optimized to enhance isotope incorporation expressing five different E5 miniprotein variants. With the currently optimized protocols we could achieve a 1.5- to 5-fold increase of yields of several miniproteins coupled to a similar magnitude of cost reduction as compared to flask labelling protocols.