Spatial structure of apamin in solution]

The calculation of the complete spatial structure of the bee venom peptide neurotoxin apamin has been carried out by means of a method elaborated earlier. It is based on the joint utilization of the molecular mechanics algorithms and NMR spectroscopy data. It was established that the molecule backbone conformation in solution may be represented as the combination of the beta-turn III (residues 2-5) and alpha-helical segment (9-18) both separated by the non-standard bend IV (5-8). The most probable system of the intramolecular hydrogen bonds in the apamin polypeptide backbone was proposed. Certain amino acid residues have been shown to be characterized by the lack of strict determination of the conformations of their side chains which may be realized in a few states providing approximately equal stabilization of the same form of the main chain. The conformational parameters of the proposed apamin structural model are appropriate to the NMR spectroscopy data derived from the literature and used in the calculations and are not contradictory to other experimental information.     

Computer modeling of promising inhibitors of the HIV-1 subtype A replication as a framework for the rational anti-AIDS drug design

The model of the structural complex of cyclophilin B belonging to the immunophilins family with the HIV-1 subtype A V3 loop presenting the principal neutralizing determinant of the virus gp120 envelope protein as well as determinants of cell tropism and syncytium formation was generated by molecular docking methods. Based on the conformational and energy characteristics of the built complex, computer-aided design of the polypeptide able to block effectively the functionally crucial V3 segments was implemented. Analysis of the results obtained in this study and literature data suggests that the generated molecule represents a promising pharmacological substance, which may be used as the basis structure for realization of the protein engineering projects aimed to develop effective drugs for anti-AIDS therapy.     

SPECT and MRT in the diagnosis of myocarditis of mild and moderate severity]

Technetium-99mWBC SPECT and contrast material enhanced MRI were used for the diagnostics of mild myocarditis. The use of labeled leukocytes and paramagnetic contrast agent allow visualizing inflammation of myocardium and assessing the efficacy of treatment. 49 patients aged 14 to 49 years old were studied. Case presentation demonstrates high information and diagnostic value of used methods.     

Geographical Ecocline Polymorphism of the Distribution of Mitochondrial Haplotypes of <Emphasis Type="Italic">Adalia bipunctata</Emphasis> Linnaeus, 1758 (Coleoptera: Coccinellidae) of Norway

Adalia bipunctata Linnaeus, 1758 is a classical object for studying polymorphism of natural populations. Application of genetic markers significantly increased the possibilities of analysis of natural populations of ladybirds and opened the possibility of studying ecological adaptations at the genetic level. We analyze the geographical distribution of the mitochondrial haplotypes of A. bipunctata, determined on the basis of the variability of the mitochondrial cox1 gene in the natural populations of Norway. Two new mitochondrial haplotypes of A. bipunctata are described. We observe a geographical cline in the haplotypes frequencies from south to north. The main haplotype H1 of the central Europe is gradually replaced in the north by the haplotype H7. A possible reason for the observed geographical cline is a selective advantage of beetles with haplotype H7 in areas with a northern coastal climate.     

Cloning and Molecular Modeling of Duodenase with Respect to Evolution of Substrate Specificity within Mammalian Serine Proteases That Have Lost a Conserved Active-Site Disulfide Bond

Mammalian serine proteases such as the chromosome 14 (Homo sapiens, Mus musculus) located granzymes, chymases, cathepsin G, and related enzymes including duodenase collectively represent a special group within the chymotrypsin family which we refer to here as "granases". Enzymes of this group have lost the ancient active-site disulfide bond Cys191-Cys220 (bovine chymotrypsinogen A numbering) which is strongly conserved in classic serine proteases such as pancreatic, blood coagulation, and fibrinolysis proteases and others (granzymes A, M, K and leukocyte elastases). We sequenced the cDNA encoding bovine (Bos taurus) duodenase, a granase with unusual dual trypsin-like and chymotrypsin-like specificity. The sequence revealed a 17-residue signal peptide and two-residue (GlyLys) activation peptide typical for granases. Production of the mature enzyme is apparently accompanied by further proteolytic processing of the C-terminal pentapeptide extension of duodenase. Similar C-terminal processing is known for another dual-specific granase, human cathepsin G. Using phylogenetic analysis based on 39 granases we retraced the evolution of residues 189 and 226 crucial for serine protease primary specificity. The analysis revealed that while there is no obvious link between mutability of residue 189 and the appearance of novel catalytic properties in granases, the mutability of residue 226 evidently gives rise to different specificity subgroups within this enzyme group. The architecture of the extended substrate-binding site of granases and structural basis of duodenase dual specificity based on molecular dynamic method are discussed. We conclude that the marked selectivity of granases that is crucial to their role as regulatory proteases has evolved through the fine-tuning of specificity at three levels--primary, secondary, and conformational.     

Electrophysiological investigation of medullary neurons connected with lateral line organs of the catfish (Ictalurus nebulosus)

Unit responses in the acoustic-lateral region of the medulla to electrical and mechanical stimulation of the lateral line organs were investigated in acute experiments on curarized catfish. Of the total number of neurons 70% possessed spontaneous activity. An electrical stimulus evoked a tonic response both in spontaneously active and in "silent" cells. Three main types of firing pattern of the neurons were distinguished: fast-adapting, slow-adapting, and grouped. As regards the relation of the neurons to polarity of the stimulus they were subdivided into two groups. The thresholds of unit responses to electrical stimulation varied considerably: from 2.5·10^–9 to 5·10^–12 A/mm². The effect of intensity of the electrical stimulation on unit responses in the medulla is analyzed. The precise dependence of on- and off-responses of each neuron on stimulus intensity of any polarity was determined. The neurons were shown to be sensitive to both electrical and mechanical stimuli. It is postulated that this phenomenon is due to convergence of impulses from electrical and mechanical receptors of the lateral line on the neurons. The properties of the central neurons are compared with those of the peripheral electroreceptor system in catfish.I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 5, No. 2, pp. 156–163, March–April, 1973.     

Local structure of cytochrome c from horse heart in solution. Conformational analysis using data of two-dimensional nuclear Overhauser effect spectroscopy]

Using the earlier suggested method the calculation of the backbone conformations of horse heart cytochrome c in oxidized (ferricytochrome c) and reduced (ferrocytochrome c) states has been performed by the two-dimensional nuclear Overhauser effect spectroscopy data. For both protein forms the secondary structure elements have been revealed and the conformations of the irregular polypeptide chain segments have been analysed. The similarity of the secondary structures of ferri- and ferrocytochrome c in solution was established from the comparison of their conformations. Small differences between the conformations of two molecule forms are shown to be localized within the polypeptide chain fragments situated in the spatial structure near the heme crevice. The comparison of the dihedral phi and psi angles in the calculated conformations of horse cytochrome C with the corresponding characteristics of X-ray structures of tuna ferri- and ferrocytochrome c made for the oxidized and reduced protein forms using the quantitative criteria testifies the similarity of their conformations in solution and crystal. In is shown that the conformational changes of the separate amino acid residues which take place as the result of the "solution-to-crystal" transition occur on the surface fragments of protein globule and do not lead to essential alterations of the secondary molecule structure.     

Changes in nitric oxide in heart of intact and sympathectomized rats of different age

Nitric oxide production in heart tissues of rats of different age in the norm and after pharmacological sympathectomy was studied by electron spin resonance spin-trapping. Rats at the age of 14, 21, 70, and 100 days were used in the experiment. The concentration of nitric oxide produced in rat heart tissues proved to considerably decrease during ontogeny. Pharmacological sympathectomy notably decreased nitric oxide production in the heart in 14-and 21-day-old rats: the nitric oxide concentration in the spin trap as well as the level of R and T conformers of hemoglobin nitrosyl complexes decreased. In 70-day-old rats, pharmacological sympathectomy had no notable effect on the level of nitric oxide-containing paramagnetic complexes.