Action of trypsin on glycinin. Mixed-type proteolysis and its kinetics; molecular mass of glycinin-T

The formation of a relatively stable high-molecular-mass product on trypsin hydrolysis of glycinin (glycinin-T) is interpreted to be a result of 'zipper' proteolysis. Evidence of parallel one-by-one degradation of glycinin occurring after the formation of glycinin-T is presented. At a relatively low concentration of the substrate, the one-by-one proteolysis proceeds as a first-order reaction. A method of determination of the changes in the molecular mass of a protein during the mixed-type proteolysis and some other parameters of this process is developed on the basis of the analysis of the proteolysis kinetics. The value of the molecular mass of glycinin-T calculated by means of this method makes up 70% of the initial molecular mass and coincides with the result of direct determination by gradient gel electrophoresis.     

Some arylamidases from vetch seeds]

Hydrolysis of L-phenylalanyl-p-nitroanilide (PPA) and glycyl-p-nitroanilide by extracts from resting vetch seeds was shown to be the effect of two different arylamidases. One of them, PPAase, was 2000-fold purified on hydroxylapatide, DEAE-cellulose and by gel filtration through Sephadex G-100. The preparation obtained was nearly homogenous chromatographycally. Molecular weight of PPAase, as shown by means of gel filtration, was 66000, K(m) was calculated to be 1.64-10-4 M. PPAase was inhibited by SH-reagents and partially by o-oxyquinoline. Some increase in the enzyme activity was observed in the presence of Ca2+, Mg2+ and Mn2+ in low concentrations. The enzyme hydrolysed amino acid arylamides with hydrophobic side groups and some dipeptides, which had at least one hydrophobic amino acid and did not contain amino acids with polar group.     

Geometrically nonlinear modelling of pre-stressed viscoelastic fibre-reinforced composites with application to arteries

Biomechanics and Modeling in Mechanobiology - Mechanical behaviour of pre-stressed fibre-reinforced composites is modelled in a geometrically exact setting. A general approach which includes two...     

Arthropod toxins inhibiting Ca2+ and Na+ channels prevent AC‐1001 H3 peptide‐induced apoptosis

Peptide toxins of arthropods are one of the potential sources of bioactive substances. Toxins are able to bind to calcium channels and block them. Ca²⁺ ions play an important role in many cell processes, in particular, in apoptosis. In this work, we study the effect of some arthropod toxins on intracellular processes associated with the induction of apoptosis. Synthetic analogs of U₅‐scytotoxin‐Sth1a, ω‐hexatoxin‐Hv1a, ω‐theraphotoxin‐Hhn2a, and μ‐agatoxin‐Aa1a toxins—inhibitors of calcium L, P, and Q channels and sodium channels were used in the study. Apoptosis was induced by AC‐1001 H3 peptide. We study the effect of toxins on the level of apoptosis, ROS, mitochondrial potential, GSH, and ATP in CHO‐K1 cells. We show that all the tested toxins are able to dose dependently block the induction of apoptosis triggered by AC‐1001 H3 and reduce the level of natural apoptosis in CHO‐K1 cells. Cell incubation with apoptosis inducer AC‐1001 H3 in the presence and absence of toxins causes an increase in the intracellular concentrations of ROS, ATP, and mitochondrial potential and decreases the GSH concentration. The present study reveals the antiapoptotic effect of a number of arthropod peptide toxins. The toxins studied can represent a novel approach used in the treatment of pathologies associated with the activation of apoptotic mechanisms.     

Comparison of the Characteristics of DC Discharges with a Liquid Anode and a Liquid Cathode over Aqueous Solutions of Zinc Nitrate

Plasma Physics Reports - Results of systematic measurements of the electric and energy characteristics of a dc gas-discharge plasma in contact with a liquid are presented. The discharge was excited...     

Description of rearrangement of distraction reclaim in the extension of the lower extremities and elimination of their deformity in patients with various etiology of shortening

The data of X-ray and computed tomographic studies of a long tube distraction reclaim were analyzed in 80 patients who had limb shortening and deformity of varying etiology. The analysis has shown that during distraction there is a positive correlation between the increase in the connective-tissue interlayer area of a distraction reclaim and the overall increase in the area of the distraction reclaim (their ratio is not greater than 30% under the optimal elongation conditions). During fixation there is a decrease in the ratio of the area of a growth zone to the total area of a reclaim and in its total density with a parallel increase in the density of cortical laminae of a newly formed bone. Complete organotypic rearrangement comes to the end in the range of 1-3 years depending on the magnitude of pathomorphological and biomechanical changes and limb function.     

Metabotropic Purinoreceptors in Rat Dorsal Horn Neurons: Predominantly Dendritic Location

Elevations of the cytosolic free Ca²⁺ concentration ([Ca²⁺] _i ) in rat dorsal horn neurons induced by addition of ATP to the medium were compared in spinal cord slices and after isolation of the neurons. In slices, application of ATP results in an increase in the [Ca²⁺] _i by 201 ± 12 nM, on the average; in a Ca²⁺ -free external solution the respective rise was 156 ± 14 nM (n = 45 of 76 examined cells), which indicate the presence of active purinergic metabotropic receptors in about 59% of the neurons. In freshly isolated neurons with absent dendrites, we found no metabotropic responses. Thus, the results confirm the conclusion on the localization of metabotropic postsynaptic purinoreceptors mostly on the dendritic tree of dorsal horn neurons.     

Legumin-like and vicilin-like seed storage proteins: Evidence for a common single-domain ancestral gene

Legumin-like 11S and vicilin-like 7S globulins are the main storage proteins of most angiosperms and gymnosperms. The subunits of the hexameric legumin are synthesized as a precursor comprising a N-terminal acidic - and a C-terminal basic -chain. The trimeric vicilin molecule consists of subunits composed of two symmetrical N- and C-terminal structural domains.In a multiple alignment we have compared the N-terminal and C-terminal domains of 11 legumns and seven vicilins of several dicot, monocot, and gymnosperm species. The comparisons using all six possible pairwise combinations reveal that the N-terminal and C-terminal domains of both protein families are similar to each other. These results together with data on the distribution of variable and conserved regions, on the positions of susceptible sites for proteolytic attack, as well as on the published 7S protein tertiary structure suggest that both protein families share a common single-domain ancestor molecule and lead to the hypothesis that a triplication event has occurred during the evolution of a putative legumin/vicilin ancestor gene.Moreover, the comparison of the intron/exon pattern reveals that at least three out of five intron positions are precisely conserved between the genes of both protein families, further supporting the idea of a common evolutionary origin of recent legumin and vicilin encoding genes. Correspondence to: H. Bäumlein