Effect of ppGpp on the accuracy of protein biosynthesis

The maintenance of accuracy in protein biosynthesis in amino acid-starved rel+ strains of Escherichia coli has been attributed to an effect of ppGpp on the accuracy of aa-tRNA selection by the ribosome. It has been determined that concentrations of ppGpp characteristic of those found in amino acid-starved cells have no effect on the rate of reaction of poly(U)-programmed ribosomes with either the cognate (Phe) or the near-cognate (Leu2) ternary complexes. Neither the rate of GTP hydrolysis, which signals selection of the ternary complex, nor the rate of peptide formation, which signals the acceptance of the aa-tRNA after proofreading, is affected by the nucleotide. The results indicate that the effect of ppGpp in maintaining the accuracy of protein biosynthesis in cells starved for an amino acid is not due to a direct effect on the rate constants for substrate selection by the ribosome.     

Calcium-dioxolene complexes: rate constants of pyrocatechol oxidation in the presence of Ca2+

The effect of calcium ions on the rate of pyrocatechol autoxidation at pH 9.0 has been studied by mathematical modeling. The effect of Ca2+ on the oxygen absorption rate has been studied, and a kinetic model has been suggested, which takes different stages of interaction of pyrocatechol and its radical form with oxygen into account. It has been shown that the prooxidant action of Ca2+ is related to an abrupt increase (approximately by three orders of magnitude) in the rate constant of comproportionation (reaction of chain branching and formation of o-semiquinonates) and a marked decrease (by two orders of magnitude, from 1.4 10(7) to 0.6 10(5) M(-1)s(-1)) in the rate constant of disproportionation of o-semiquinones. The system can be used as a model for studying the prooxidant action of calcium ions.     

Effect of sucrose on the optical properties of dipalmitoylphosphatidylcholine

The gel to liquid crystal phase transition of dipalmitoylphosphatidylcholine (DPPC) has been followed by the change in absorbance at 400 nm; this change is due to the change in lipid light scattering properties during the transition. The effect of sucrose on the change in absorbance during the transition of DPPC has been investigated. It has been shown that the presence of sucrose or glycerol in the multilamellar liposome suspension increases the change in absorbance due to the main transition, decreases the total absorbance, and decreases the change in absorbance due to the pretransition. This effect of sucrose and glycerol is shown to be an optical effect which is correlated with solvent index of refraction.     

Effect of a hypertonic mannitol solution on the peri-infarct area of an isolated frog heart

The influence of the hypertonic (3%) solution of mannitol on the process of the excitability recovery (in the near necrotic zone) appeared as a result of the necroses tissue effect on the ventricle has been investigated in the experiments on the isolated frog's heart. The analogous perfusion was conducted by Ringer solution in the control experiments. The process of isopotential phase S-T recovery was practically the same at heart perfusion with Ringer solution or with mannitol. Simultaneous registration of ventricular mechanogram showed the pronounced negative inotropic effect of mannitol. It is suggested that the positive effect of mannitol which has been noted under clinic conditions during the treatment of ischemic disease, doesn't connect with the effect of hypertonic solution on the cells excitability of periinfarcted (near necrotic) zone.     

Effect of ionizing radiation on the structure of human serum albumin. Effect of irradiation on the content of free amino groups and on the ability of HSA to be digested by proteinases

The effect of gamma-quanta on human serum albumin (HSA) solutions (1.6 mg/ml, borate buffer pH 7.45) in the air has been investigated. Using 2,4,6-trinitro-benzene-sulfonic acid it has been shown that 2-22 kGy radiation reduces the free amino groups content of HSA and increases its resistance to the hydrolytic effect of trypsin and pronase which is not influenced by the postirradiation exposure to heat. It is concluded that epsilon-NH2-groups of lysine residues are modified and firm cross-links are formed in HSA under the effect of radiation.     

State of differentiation of bovine epithelial lens cells in vitro. Relationship between the variation of the cell shape and the synthesis of crystallins

Correlations between cell morphology and the expression of specific proteins (crystallins) have been investigated. Two different culture conditions have been chosen which keep bovine epithelial lens cells (BEL cells) in a monolayer of homogeneous epithelioid cells: (1) bovine retinal extract (EDGF) supplemented medium; (2) extracellular matrix (ECM) provided by corneal endothelial cells in standard medium has been compared to previous results obtained with BEL cells cultivated on plastic (Simonneau et al., 1983). Variations of the cell shape had no effect upon crystallin synthesis.     

Lipid intermediates in the biosynthesis of the wall teichoic acid in Staphylococcus lactis I3

1. Particulate enzyme systems have been prepared from Staphylococcus lactis I3 which effect the synthesis of wall teichoic acid (a polymer containing a repeating unit in which d-glycerol 1-phosphate is attached to the 4-position on N-acetylglucosamine 1-phosphate) from the nucleotide precursors CDP-glycerol and UDP-N-acetylglucosamine. By using nucleotides labelled with (32)P and (14)C it has been shown that the synthesis proceeds via lipid intermediates. 2. Two intermediates have been found. In one of these N-acetylglucosamine 1-phosphate is present, whereas in the other the repeating unit of the teichoic acid occurs. 3. The simultaneous formation of the teichoic acid, a poly-(N-acetylglucosamine 1-phosphate) and an unidentified lipid, together with the poor ability of most particulate systems to synthesize polymer and the instability of the lipid intermediates themselves, have interfered with pulse-labelling experiments. Nevertheless, the biosynthetic sequence has been elucidated. It is concluded that the intermediates are derivatives of undecaprenol phosphate.     

Symphoria: the success of modeling the active site function of oxo-molybdoenzymes

The success of modeling the active site function of oxomolybdoenzymes have been claimed generally on the basis of reactivity of the synthetic analogues towards PPh(3) or DMSO (dimethyl sulfoxide). Here it has been shown that the success of modeling the active site function of these enzymes may not be determined by the ability of a model to undergo oxotransfer with PPh(3) or DMSO (except for the modeling of DMSO reductase) and one should adhere to the criteria accepted by the bioinorganic community. A critical evaluation of two of those criteria which requires a synthetic analogue (a) should react with the enzyme substrate (b) should follow the same rate law as does the enzyme, has been presented in this paper. We have shown that the fulfillment of criterion (b) and the inhibition phenomena to that effect both are dictated by symphoria (from sympherin in Greek: the bringing together of reactants into the proper spatial relationship) on the basis of kinetic studies of the reactivity of enzyme substrate the HSO(3)(-) and its analogues (anions of oxyacids of phosphorous) towards a functional model sulfite oxidase [Bu(4)N](2)[Mo(VI)O(2)(mnt)(2)] (mnt(2-)=1,2-dicyanoethylenedithiolate) but with the caveat that the mechanistic inference drawn from such studies may not be the same as in the case of native enzyme. In view of this ambiguity it has been pointed out that the fulfillment of this criterion is not a definitive conclusion towards our understanding of the structure-function relationship of an enzyme and, therefore, the criterion of a 'structural analogue' and 'functional analogue' have been revised subject to an amendment of criterion (a) to include substrate analogues. It has also been shown for the first time on the basis of kinetic studies that the effect of medium can lead to substrate - inhibitor type dualism and hence the effect of medium is also a factor that can play a key role for the success of modeling the active site function of an enzyme. Here we also provide the details of the inhibition mechanisms proposed in our earlier report with an indirect proof to that effect.     

A photosynergistic lethal effect in the irradiation of yeasts with long-wave ultraviolet and visible light

A lethal synergistic effect which is expressed as the nonadditive summing of the damaging effect of each irradiation separately has been found during the investigation of combined action of longwave ultraviolet (UV) rays (337 nm or 365 nm) and visible light (400-600 nm) on the yeast cells. Based on the data on different mechanisms of lethal effect of longwave UV and visible light, it has been suggested that the basis of the photosynergistic effect is the mutual intensification of the photo-destructive processes occurring in different intracellular structures and processes induced by different endogenous sensitizers.     

Role of the oncogene in the mutagenic activity of bovine adenovirus type 3

The mutagenic and carcinogenic effect of two EcoRI-fragments of bovine adenovirus type 3 (BAV-3) DNA inserted into pBR325 has been studied. The C fragment (located between 3,6 and 19,7 map units) contains the viral oncogene, the C fragment (between 44,3 and 63,7 map units) displays no transforming activity. It has been established that oncogene BAV-3 statistically true increases the yield of mutants resistant to 6-mercaptopurine (6MP) in Chinese hamster cells. The C fragment, pBR325 without viral sequences and DNA fragments of different molecular weights from normal Syrian hamster cells have no mutagenic effect. The control over tumor formation in syngenic mice after injection of C3H10T 1/2 and D. C fragments and pBR325 treatment exposed a parallelism between the mutagenic and transforming effect. The study of the combined effect of viral DNA fragments and the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) which increases the transforming activity of different carcinogens, shows that the promoter increases the frequency of mutants after viral oncogene treatment and does not induce mutagenic activity of those types of DNA which are unable to transform the cells.     

The effect of anthropogenic acidification on the hydrofauna of the lakes of the West Tatra Mountains (Slovakia)

Fourteen West Tatra lakes were studied, of which one could be considered to be recently anthropogenically acidified and eight others classified as acidification-endangered. In the anthropogenically acidified lake, the zooplankton assemblage has been substantially altered (three mountain-lake crustacean species have been eliminated). Several littoral macrobenthic species sensitive to acidification have either been eliminated from the acidified and acidification-endangered lakes or occur only sporadically. The effect of acidification has so far not been observed on the benthic fauna of the lakes medial which is probably due to the higher pH below the surface of sediment. In comparison with the High Tatra, acid depositions have had a less pronounced effect on the lakes of the West Tatra.     

Absorption of light in photoreceptors: Effect of waveguiding property

The effect of waveguiding property (i.e., the intensity distribution) of the photoreceptor on the number of photons absorbed in a photoreceptor has been studied. It has been found that the effect is significant only for large values of the exposure and the maximum effect is less than 11% in the case of human rod photoreceptor. In the analysis, the funnelling effect, which follows from the coupling between the interior and exterior fields, has not been considered.Work partially supported by the Department of Science and Technology (India)B. D. Gupta is associated with the School of Bioscience Studies     

Error minimization explains the codon usage of highly expressed genes in Escherichia coli

Different organisms use synonymous codons with different preferences. Several measures have been introduced to compute the extent of codon usage bias within a gene or genome, among which the codon adaptation index (CAI) has been shown to be well correlated with mRNA levels of Escherichia coli. In this work an error adaptation index (eAI) is introduced, which estimates the level at which a gene can tolerate the effects of mistranslations. It is shown that the eAI has a strong correlation with CAI, as well as with mRNA levels, which suggests that the codons of highly expressed genes are selected so that mistranslation would have the minimum possible effect on the structure and function of the related proteins.     

Intracardiac neurogenic regulation of the rate of excitation conduction in the atrioventricular area of the heart. I. Dependence of the effect on the moment of stimulus application in spontaneous and artificially stabilized rhythms of the heart beat

The influence of the intracardiac reflectory system on the atrioventricular propagation in the experiments on the isolated frog's heart has been investigated. The ability of the intracardiac reflectory cholinergic activation to accelerate or reduce the atrioventricular propagation was confirmed. Both effects remain on the background of the artificially stabilized heart rhythm. The dromotropic effect direction was shown not to be connected with the electrocardiogram (ECG) phase at which the stimulus has been given; the positive or negative dromotropic effect can appear giving the stimulus at the same phase of the ECG. But there is a strong correlation between the moment of the stimulation and the dromotropic effect intensity. The most high degree of the acceleration and inhibition appears when the stimulus is given near the P component of the ECG.     

The photosensitivity of the malate oxidase system of a pigmented strain and a carotenoidles mutant of Sarcina lutea (Micrococcus luteus)

The effect of white light on the malate oxidase of Sarcina lutea (Micrococcus luteus) membranes has been examined using a carotenoid-containing and a carotenoidless mutant. At least three photosensitive sites have been detected. Two of these are associated with the malate dehydrogenase complex (malate-menaquinone reductase) and are unaffected by membrane carotenoid. A third site which has been detected beyond the dehydrogenase complex, is protected by carotenoid since it can only be demonstrated in carotenoidless systems. A repair mechanism has been found for one of the two sites in the dehydrogenase complex.     

Stabilization of yeast hexokinase A by polyol osmolytes: correlation with the physicochemical properties of aqueous solutions

Osmolytes of the polyol series are known to accumulate in biological systems under stress and stabilize the structures of a wide variety of proteins. While increased surface tension of aqueous solutions has been considered an important factor in protein stabilization effect, glycerol is an exception, lowering the surface tension of water. To clarify this anomalous effect, the effect of a series of polyols on the thermal stability of a highly thermolabile two domain protein yeast hexokinase A has been investigated by differential scanning calorimetry and by monitoring loss in the biological activity of the enzyme as a function of time. A larger increase in the T(m) of domain 1 compared with that of domain 2, varying linearly with the number of hydroxyl groups in polyols, has been observed, sorbitol being the best stabilizer against both thermal as well as urea denaturation. Polyols help retain the activity of the enzyme considerably and a good correlation of the increase in T(m) (DeltaT(m)) and the retention of activity with the increase in the surface tension of polyol solutions, except glycerol, which breaks this trend, has been observed. However, the DeltaT(m) values show a linear correlation with apparent molal heat capacity and volume of aqueous polyol solutions including glycerol. These results suggest that while bulk solution properties contribute significantly to protein stabilization, interfacial properties are not always a good indicator of the stabilizing effect. A subtle balance of various weak binding and exclusion effects of the osmolytes mediated by water further regulates the stabilizing effect. Understanding these aspects is critical in the rational design of stable protein formulations.     

Cation-dependence of the self-association behavior of guanylyl-(3'-5')-guanosine.

The aggregation behavior of guanylyl-(3'-5')-guanosine, GpG, in the form of the tetramethylammonium (TMA), Li, Na, and K salts in aqueous solution has been investigated by NMR and FTIR techniques. The salts were prepared by a cation-exchange method. The ability of the cations to induce aggregate formation is TMA+ < Li+ < Na+ < K+, where TMA+ has only a weakly promoting action and K+ has a very strong effect. Three types of aggregates have been observed: (a) small aggregates which are in rapid exchange with respect to the NMR time scale; (b) intermediate-sized aggregates which are slow to exchange; (c) very large aggregates which can only be observed by FTIR. In all cases the aggregated species are held together by base stacking and guanine-guanine hydrogen bonding. A stoichiometry of 2 GpG per K+ has been determined by a 1H NMR titration of TMAGpG with KCl. Models have been proposed for the various-sized species. These include stacked dimers, stacked tetramers (similar to G-tetrads), and species in which K+ ion bridges between phosphates in separate tetramers.