Study of the pyruvate dehydrogenase complex by circular dichroism]

A comparative study of the pyruvate dehydrogenase complex and its pyruvate dehydrogenase component was carried out by using the circular dichroism method. It was found that the spectral properties of the pyruvate dehydrogenase complex are determined by those of its first component: i) the spectrum of the thiamine pyrophosphate-free pyruvate dehydrogenase complex displayed the main characteristics of the pyruvate dehydrogenase component; ii) the appearance of the charge transfer complex band during thiamine pyrophosphate saturation was revealed for the both proteins; iii) in both cases the charge transfer complex band disappeared after the interaction of the holoform with pyruvate and reappeared after the addition of dithiothreitol used as a deacetylating reagent. Coenzyme A in the same reaction selectively deacetylated the pyruvate dehydrogenase complex (but not its pyruvate dehydrogenase component). The spectral dynamics of pyruvate dehydrogenase reflects the functional changes in the enzyme active centers during the catalytic act. The similarity of the spectral behaviour of pyruvate dehydrogenase within the complex structure and in the isolated state provides support for the earlier proposed mechanism of the pyruvate dehydrogenase action and ensures a methodological basis for its direct investigation within the complex structure.     

Metabolic level and the function of the peripheral catecholaminergic systems in immobilization hypothermia in rats

Metabolic peculiarities were studied on the model of prolonged immobilization hypothermia in rats (body temperature +20 degrees C for 24 h). Stress reactions and the state of peripheral catecholaminergic systems were compared in hypo- and normothermia. A direct correlation was established between the intensity of metabolism and the mediator activity in adrenergic nerve structures.     

Mathematical model of carbohydrate energy metabolism. Interaction between glycolysis, the Krebs cycle and the H-transporting shuttles at varying ATPase load

A simple mathematical model for carbohydrate energy metabolism based on the stoichiometic structure of glycolysis, the Krebs cycle and oxidative phosphorylation is proposed. The only allosteric regulation involved in the model is phosphofructokinase activation by AMP. Simple as it is, the model can explain the following properties of carbohydrate metabolism: a drastic rise of the rate of glucose consumption during transition to a higher level of ATPase load; stabilization of ATP and an increase of the steady state rates of glycolysis and oxidation of cytoplasmic NADH by the H-transporting shuttles and of pyruvate in the Krebs cycle with increasing rate of the ATPase load; activation of glycolysis and a decrease of the rate of oxidative phosphorylation following an inhibition of the H-transporting shuttles. The mechanisms of the coordinated changes in the steady state rates of glycolysis, the H-transporting shuttles and the Krebs cycle at varying ATPase load in the cell are discussed.     

C-terminal fragment of ribosomal protein S15 is located at the decoding site of the human ribosome

Protein S15 is a characteristic component of the mammalian 80S ribosome that neighbors mRNA codon at the decoding site and the downstream triplets. In this study we determined S15 protein fragments located close to mRNA positions +4 to +12 with respect to the first nucleotide of the P site codon on the human ribosome. For cross-linking to ribosomal protein S15, a set of mRNA was used that contained triplet UUU/UUC at the 5'-termini and a perfluorophenyl azide-modified uridine in position 3' of this triplet. The locations of mRNA analogues on the ribosome were governed by tRNAPhe cognate to the UUU/UUC triplet targeted to the P site. Cross-linked S15 protein was isolated from the irradiated with mild UV light complexes of 80S ribosomes with tRNAPhe and mRNA analogues with subsequent cleavage with CNBr that splits polypeptide chain after methionines. Analysis of modified oligopeptides resulted from the cleavage revealed that in all cases cross-linking site was located in C-terminal fragment 111-145 of protein S15 indicating that this fragment is involved in formation of decoding site of the eukaryotic ribosome.     

Factors contributing to preservation of Vibrio cholerae in water reservoirs

The summarized data of literature concerning the survival of V. cholerae in the environment and the influence of abiotic and biotic factors on this process are presented. These data make it possible to regard cholera as sapronosis and to form an idea of the role of factors contributing to the survival of V. cholerae in the environment and to its spread among human population.     

Isolation and characteristics of a plasmid from the thermotolerant strain of Bacillus licheniformis

A 8.3 kb cryptic plasmid was isolated from the thermotolerant strain of Bacillus licheniformis 28KA and designated pLT83. The replicative (rep) region was localized on the plasmid map. The pLT83 plasmid labelled in vitro with an antibiotic resistance determinant is able to replicate in B. subtilis cells. The pLT83 plasmid replicates stably in B. licheniformis strain at higher temperatures (37-60 degrees C) than in B. subtilis cells (37-50 degrees C). The plasmid and its derivatives may be used as vectors for gene cloning in B. subtilis and B. licheniformis cells.     

Cloning of the genes controlling the biosynthesis of bacitracin in Bacillus licheniformis

The DNA fragment from bacitracin-producing Bacillus licheniformis strain is cloned on pMX39 vector plasmid in Bacillus subtilis cells. Bacillus subtilis cells carrying the cloned fragment inhibit the growth of bacitracin-sensitive tester strain. The observed inhibition of growth is due to the production by Bacillus subtilis of bacteriocin substance that is identified as bacitracin by TLC-chromatography. In contrast to the data published earlier it is shown that Bacillus subtilis can in fact produce the small amounts of bacitracin. Introduction of the cloned Bacillus licheniformis DNA into Bacillus subtilis cells stimulates this bacitracin production. The restriction site map of the Bacillus licheniformis chromosomal region bearing the cloned fragment is constructed.     

Bovine papillomavirus type 1 infection is mediated by SNARE syntaxin 18

Events that lead to viral infections include the binding of the virus to the target cells, internalization of the virus into the cells, and the ability of the viral genome to be expressed. These steps are mediated by cellular and viral proteins and are temporally regulated. The papillomavirus capsid consists of two virally encoded capsid proteins, L1 and L2. Much is known about the role of the major capsid protein L1 compared to what is known of the role of the L2 protein. We identified the interaction of the L2 protein with SNARE protein syntaxin 18, which mediates the trafficking of vesicles and their cargo between the endoplasmic reticulum, the cis-Golgi compartment, and possibly the plasma membrane. Mutations of L2 residues 41 to 44 prevented the interaction of L2 protein with syntaxin 18 in cotransfection experiments and resulted in noninfectious pseudovirions. In this paper, we describe that syntaxin 18 colocalizes with infectious bovine papillomavirus type 1 (BPV1) pseudovirions during infection but does not colocalize with the noninfectious BPV1 pseudovirions made with an L2 mutant at residues 41 to 44. We show that an antibody against BPV1 L2 residues 36 to 49 (alpha L2 36-49) binds to in vitro-generated BPV1 pseudoviral capsids and does not coimmunoprecipitate syntaxin 18- and BPV1 L2-transfected proteins. alpha L2 36-49 was able to partially or completely neutralize infection of BPV1 pseudovirions and genuine virions. These results support the dependence of syntaxin 18 during BPV1 infection and the ability to interfere with infection by targeting the L2-syntaxin 18 interaction and further define the infectious route of BPV1 mediated by the L2 protein.     

Antibiotic sensitivity of Vibrio cholerae 01 isolated from environmental objects]

The sensitivity of 252 Vibrio cholerae-O1 strains isolated from environmental objects to antibiotics of various groups was assayed by the method of serial dilutions on solid media. The biological characteristics of the isolates are presented. The Vibrio cholerae isolates with serological variation were the most frequent (36.6 per cent), so are the cultures detected by their sensitivity to the specific phages (87.5 per cent). It was found that changes in some biological properties of the strains did not coincide with the changes in the antibiotic sensitivity. The isolates were highly sensitive to tetracycline, chloramphenicol, gentamicin, erythromycin and rifampicin and less sensitive to novobiocin and the other aminoglycosides. The sensitivity to the beta-lactams was the lowest. The resistance determinants were detected in single strains (6.3 per cent), the kanamycin and novobiocin resistance determinants being detected in 15 out of the 16 strains tested. The study showed that the cultures of Vibrio cholerae-O1 isolated from the environmental objects generally preserved their sensitivity to the diverse group antibiotics.