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31.
We present the computer program hybridlab 1.0 for simulating intraspecific hybrids from population samples of nuclear genetic markers such as microsatellites, allozymes or SNPs (single nucleotide polymorphisms). The program generates a user‐specified number of multilocus F1 hybrid genotypes between any pair of potentially hybridizing populations included in a standard input‐file of multilocus genotypes for population genetic analysis. This simple, user‐friendly program has a wide range of applications for studying natural and artificial hybridization; in particular, for evaluating the statistical power for individual assignment of parental and hybrid individuals. An example of application for Atlantic cod populations is given. 相似文献
32.
Valentin V. Makarov Eugeny V. Skurat Pavel I. Semenyuk Dmitry A. Abashkin Natalya O. Kalinina Alexsandr M. Arutyunyan Andrey G. Solovyev Eugeny N. Dobrov 《PloS one》2013,8(4)
Virions of Barley stripe mosaic virus (BSMV) were neglected for more than thirty years after their basic properties were determined. In this paper, the physicochemical characteristics of BSMV virions and virion-derived viral capsid protein (CP) were analyzed, namely, the absorption and intrinsic fluorescence spectra, circular dichroism spectra, differential scanning calorimetry curves, and size distributions by dynamic laser light scattering. The structural properties of BSMV virions proved to be intermediate between those of Tobacco mosaic virus (TMV), a well-characterized virus with rigid rod-shaped virions, and flexuous filamentous plant viruses. The BSMV virions were found to be considerably more labile than expected from their rod-like morphology and a distant sequence relation of the BSMV and TMV CPs. The circular dichroism spectra of BSMV CP subunits incorporated into the virions, but not subunits of free CP, demonstrated a significant proportion of beta-structure elements, which were proposed to be localized mostly in the protein regions exposed on the virion outer surface. These beta-structure elements likely formed during virion assembly can comprise the N- and C-terminal protein regions unstructured in the non-virion CP and can mediate inter-subunit interactions. Based on computer-assisted structure modeling, a model for BSMV CP subunit structural fold compliant with the available experimental data was proposed. 相似文献
33.
Pavel I. Semenyuk Vladimir I. Muronetz Thomas Haertlé Vladimir A. Izumrudov 《Biochimica et Biophysica Acta (BBA)/General Subjects》2013
Background
It is well documented that poly(sulfate) and poly(sulfonate) anions suppress protein thermal aggregation much more efficiently than poly(carboxylic) anions, but as a rule, they denature protein molecules. In this work, a polymer of different nature, i.e. poly(phosphate) anion (PP) was used to elucidate the influence of phosphate groups on stability and thermal aggregation of the model enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH).Methods
Isothermal titration calorimetry and differential scanning calorimetry were used for studying the protein–polyanion interactions and the influence of bound polyanions on the protein structure. The enzymatic activity of GAPDH and size of the complexes were measured. The aggregation level was determined from the turbidity.Results
Highly polymerized PP chains were able to suppress the aggregation completely, but at significantly higher concentrations as compared with poly(styrenesulfonate) (PSS) or dextran sulfate chains of the same degree of polymerization. The effect of PP on the enzyme structure and activity was much gentler as opposed to the binding of dextran sulfate or, especially, PSS that denatured GAPDH molecules with the highest efficacy caused by short PSS chains. These findings agreed well with the enhanced affinity of polysulfoanions to GAPDH.Conclusions
The revealed trends might help to illuminate the mechanism of control of proteins functionalities by insertion of charged groups of different nature through posttranslational modifications.General significance
Practical implementation of the results could be the use of PP chains as promising tools to suppress the proteins aggregation without noticeable loss in the enzymatic activity. 相似文献34.
Anna S. Chalova Maria V. Sudnitsyna Pavel I. Semenyuk Victor N. Orlov Nikolai B. Gusev 《Cell stress & chaperones》2014,19(6):963-972
Temperature-induced conformational changes of reduced and oxidized HspB1 crosslinked by disulfide bond between single Cys137 of neighboring monomers were analyzed by means of different techniques. Heating of reduced HspB1 was accompanied by irreversible changes of Trp fluorescence, whereas oxidized HspB1 underwent completely reversible changes of fluorescence. Increase of the temperature in the range of 20–70 °C was accompanied by self-association of both reduced and oxidized protein. Further increase of the temperature led to formation of heterogeneous mixture of large self-associated complexes of reduced HspB1 and to formation of smaller and less heterogeneous complexes of oxidized HspB1. Heat-induced changes of oligomeric state of reduced HspB1 were only partially reversible, whereas the corresponding changes of oligomeric state of oxidized HspB1 were almost completely reversible. Oxidation resulted in decrease of chaperone-like activity of HspB1. It is concluded that oxidative stress, inducing formation of disulfide bond, can affect stability and conformational mobility of human HspB1. 相似文献
35.
LP Kurochkina PI Semenyuk VN Orlov J Robben NN Sykilinda VV Mesyanzhinov 《Journal of virology》2012,86(18):10103-10111
Chaperonins promote protein folding in vivo and are ubiquitously found in bacteria, archaea, and eukaryotes. The first viral chaperonin GroEL ortholog, gene product 146 (gp146), whose gene was earlier identified in the genome of bacteriophage EL, has been shown to be synthesized during phage propagation in Pseudomonas aeruginosa cells. The recombinant gp146 has been expressed in Escherichia coli and characterized by different physicochemical methods for the first time. Using serum against the recombinant protein, gp146's native substrate, the phage endolysin gp188, has been immunoprecipitated from the lysate of EL-infected bacteria and identified by mass spectrometry. In vitro experiments have shown that gp146 has a protective effect against endolysin thermal inactivation and aggregation, providing evidence of its chaperonin function. The phage chaperonin has been found to have the architecture and some properties similar to those of GroEL but not to require cochaperonin for its functional activity. 相似文献
36.
EG Smirnova GS Muromtsev AV Osipenko EE Khavkin LS Yaguzhinsky 《Biochemistry. Biokhimii?a》1998,63(9):1021-1028
Activation of highly specific biochemical processes by simple chemical agents is demonstrated for morphogenesis (anlage and development of female gametophyte in cereal) and mitosis (in cell cultures and animal and plant tissues). The effects of these agents are tissue-specific. Structure--activity relationship is analyzed in this group of compounds. Thus, the phenomenon reveals the exact pathways of the influence of allelopathic and anthropogenic chemical agents on evolution of plant biocenoses. 相似文献
37.
Sialylation is a biosynthetic process occurring in the trans compartments
of the Golgi apparatus. Corresponding evidence is based on localization and
biochemical studies of alpha2, 6(N)-sialyltransferase (ST6Gal I) as
previously reported. Here we describe generation and characterization of
polyclonal antibodies to recombinant rat alpha2,3(N)-sialyltransferase
(ST3Gal III) expressed as a soluble enzyme in Sf9 cells or as a
beta-galactosidase-human-ST3Gal III fusion- protein from E.coli ,
respectively. These antibodies were used to localize ST3Gal III by
immunofluorescence in various cell lines and rat kidney tissue sections. In
transiently transfected COS cells the antibodies directed to soluble
sialyltransferase or the sialyltransferase portion of the fusion-protein
only recognized the recombinant antigen retained in the endoplasmic
reticulum. However, an antibody fraction crossreactive with
beta-galactosidase recognized natively expressed ST3Gal III which was found
to be colocalized with beta1, 4-galactosyltransferase in the Golgi
apparatus of several cultured cell lines. Antibodies affinity purified on
the beta- galactosidase-ST3Gal III fusion-protein column derived from both
antisera have then been used to localize the enzyme in perfusion-fixed rat
kidney sections. We found strong staining of the Golgi apparatus of tubular
epithelia and a brush-border-associated staining which colocalized with
cytochemical staining of the H+ATPase. This subcellular localization was
not observed for ST6Gal I which localized to the Golgi apparatus. These
data show colocalization in the Golgi apparatus and different post-Golgi
distributions of the two sialyltransferases.
相似文献
38.
Semenyuk EG Stremovskiy OA Edelweiss EF Shirshikova OV Balandin TG Buryanov YI Deyev SM 《Biochimie》2007,89(1):31-38
We successfully cloned and expressed a single-chain antibody (425scFv), that is directed to human epidermal growth factor receptor HER1 (EGFR) in transgenic tobacco plants as a fusion with bacterial barstar gene (425scFv-barstar). Plant-produced recombinant 425scFv-barstar was recovered using barstar-barnase system. Based on barstar-barnase affinity, during purification of the plant-produced 425scFv-barstar, we generated bispecific scFv-antibody heterodimers from individual single-chain fragments initially produced in different host systems with binding activity to both HER1 and HER2/neu tumor antigens. We demonstrated by flow cytometry and indirect immunofluorescent microscopy that both the components of heterodimer retain its specific cell-binding activity. 相似文献
39.
40.
Remo Freimann Helmut Bürgmann Stuart EG Findlay Christopher T Robinson 《The ISME journal》2013,7(12):2361-2373
Glaciated alpine floodplains are responding quickly to climate change through shrinking ice masses. Given the expected future changes in their physicochemical environment, we anticipated variable shifts in structure and ecosystem functioning of hyporheic microbial communities in proglacial alpine streams, depending on present community characteristics and landscape structures. We examined microbial structure and functioning during different hydrologic periods in glacial (kryal) streams and, as contrasting systems, groundwater-fed (krenal) streams. Three catchments were chosen to cover an array of landscape features, including interconnected lakes, differences in local geology and degree of deglaciation. Community structure was assessed by automated ribosomal intergenic spacer analysis and microbial function by potential enzyme activities. We found each catchment to contain a distinct bacterial community structure and different degrees of separation in structure and functioning that were linked to the physicochemical properties of the waters within each catchment. Bacterial communities showed high functional plasticity, although achieved by different strategies in each system. Typical kryal communities showed a strong linkage of structure and function that indicated a major prevalence of specialists, whereas krenal sediments were dominated by generalists. With the rapid retreat of glaciers and therefore altered ecohydrological characteristics, lotic microbial structure and functioning are likely to change substantially in proglacial floodplains in the future. The trajectory of these changes will vary depending on contemporary bacterial community characteristics and landscape structures that ultimately determine the sustainability of ecosystem functioning. 相似文献