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Biomedical subjects

A Donchev

Publications and source records attributed to A Donchev.

11 recordsLinked to original sources

Comparative study of the stability of poplar plastocyanin isoforms.

The stability of the two isoforms of poplar plastocyanin (PCa and PCb) was studied with differential scanning calorimetry (DSC) technique. It was shown that the thermal unfolding of both isoforms is an irreversible process with two endothermic and one exothermic peaks. The melting temperature of PCb was found to be 1.3+/-0.2 K degrees higher than of PCa, which indicates that PCb is more stable. The enthalpy of unfolding was estimated from the heat capacity curves and was found to be significantly higher for PCb at salt concentration I=0.1 M. In addition, PCb unfolding enthalpy and melting temperature are much more sensitive to the changes in the salt concentration as found in the experiments done at different ionic strength. The experiments were complemented with numerical calculations. The salt effect on the stability was modeled using the X-ray structure of PCa and a homology modeled structure of PCb. It was found, in agreement with the experimental data, that the stability of PCb changes by 4.7 kJ more than PCa, as the salt concentration increases from zero to 0.1 M. Thus, the differences in only 12 amino acid positions between "a" and "b" isoforms result in a measurable difference in the folding enthalpy and a significant difference in the salt dependence. The optimization of the electrostatic energies of PCa and PCb were studied and it was shown that PCb is better electrostatically optimized.

Plastocyanin↗

Experimental and numerical study of the poplar plastocyanin isoforms using Tyr as a probe for electrostatic similarity and dissimilarity.

A detailed study of the tyrosine spectral characteristics was carried out in a broad range of pHs for both isoforms of plastocyanin from poplar. It was found that Tyr 80 is always protonated while Tyr 83 can form a tirosinate at high pHs. The pK(a) of Tyr 83 is practically identical in plastocyanin a and b, but the quenching of its spectrum is different in the isoforms. This provides insights that the acidic patches surrounding Tyr 83 have different electrostatic properties in plastocyanin a and b. The protonation states and the electrostatic interactions were numerically modeled on the existing plastocyanin a structure and on a homology model of plastocyanin b. The results of numerical calculations agree with the experimental findings and identify several differences in the titration behavior of the acidic patches. The difference of the tyrosine quenching pH profiles of the isoforms is rationalized by the differences in the calculated pK(a)'s of amino acids in the neighboring acidic clusters.

Acrylamide↗

Extra- and intra-cellular lytic effects of Cytophaga sp. LR2 on the red microalgae Rhodella reticulata.

AIMS: To evaluate the lytic activities of crude enzymes from Cytophaga sp. LR2 on Rhodella reticulata cells and isolated algal polysaccharide. METHODS AND RESULTS: The Cytophaga compartment was separated after centrifugation in a cell suspension for 30 min at 18,000 g. The extracellular enzyme was obtained from the supernatant and the intracellular from the pelleted cells after sonication and removal of debris. Algal cells were incubated with extra- or intracellular preparations and sowed onto agar medium. The suppressive effect of the extracellular enzyme on colony-forming units was found to be almost twice as high. The result was still more pronounced when treated cells had been shocked osmotically before seeding. Saccharolytic activity was evaluated by changes in the reducing sugars in the media. Concerning isolated algal polysaccharide, the reducing power of the two bacterial preparates was relatively low. A combined fraction showed the highest lytic activity. Using native and SDS electrophoresis some relation between the prevalence of the extra and intracellular protein patterns was registered. Two of the common components' molecular weight masses of 50 and 21 kDa were found to be reproducible in native- and SDS-containing gel. CONCLUSIONS: Cytophaga sp. LR2 produce extra- and intracellular enzymes active in destroying Rhodella cultures. The agents excreted in the medium are more effective.We suppose that two or three different classes of enzymes are involved in the lysis process. The comparative electrophoresis in this case shows the protein components with predictable functions. SIGNIFICANCE AND IMPACT OF THE STUDY: Combining different simple and reproducible approaches to identify the lytic capability of Cytophaga sp. LR2 on R. reticulata.

Cell Count↗

[Ga18(SitBu3)8] and [Ga22(SitBu3)8]--syntheses and structural characterization of novel gallium cluster compounds.

The novel neutral gallium cluster compounds [Ga18R*8] (1) and [Ga22R*8] (2) are obtained by warming up a metastable solution of gallium(I) bromide in THF/C6H5CH3 after addition of equimolar amounts of supersilyl sodium NaR* from -78 degrees C to room temperature (R* = SitBu3 = supersilyl). From X-ray structure analyses, the observed arrangements of the 18 and 22 Ga atoms in 1 and 2, respectively, are comparable with an 18 atom section of the beta-Ga modification, or show at least some kind of relationship to a 22 atom section of the Ga-III modification. This allows a description of both the clusters as metalloid. The topology of the atoms in 2 is also well explained by the Wade-Mingos rules as an eightfold capped closo-Ga14 cluster, whereby the Ga atoms of Ga14 occupy the center and the corners of a cuboctahedron with one Ga3 face replaced by a Ga4 face. Some concepts are presented about the formation mechanism, the cluster growth, and the metalloid character of the two Ga cluster compounds.

Journal Article↗