Topography of nucleic acid helices in solutions. 3. Interactions of spermine and spermidine derivatives with polyadenylic-polyuridylic and polyinosinic-polycytidylic acid helices

The synthesis of spermine derivatives (II), \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm R}_1 {\rm R}_{\rm 2} {\rm R}_{\rm 3} \mathop {\rm N}\limits^ + \left( {{\rm CH}_2 } \right)_3 \mathop {\rm N}\limits^ + {\rm R}_{\rm 1} {\rm R}_{\rm 2} \left( {{\rm CH}_2 } \right)_2 ]_2 \cdot 4{\rm X}^ - $\end{document}, and spermidine derivatives (III), \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm R}_1 {\rm R}_{\rm 2} {\rm R}_{\rm 3} \mathop {\rm N}\limits^ + \left( {{\rm CH}_2 } \right)_4 \mathop {\rm N}\limits^ + {\rm R}_{\rm 1} {\rm R}_{\rm 2} \left( {{\rm CH}_2 } \right)_3 \mathop {\rm N}\limits^ + {\rm R}_{\rm 1} {\rm R}_{\rm 2} {\rm R}_3 \cdot 3{\rm X}^ - $\end{document}, are reported. The effects of these salts on the helix–coil transition of rA–rU and rI–rC helices were examined. Increasing the size of the hydrophobic substituents, R¹, R², and R³ lowers the degree of stabilization of the helical structure. The disproportionation reaction, 2rA–rU→rA–rU₂ + rA occurs readily with salts II and III, especially when the substituents, R₁, R₂, and R₃ are small, i.e., H or Me. Spermine is found to stabilize the rA–rU² and rI–rC helices to approximately the same extent; however, large differences between the degree of stabilization of rA–rU₂ and rI-rC helices are observed when the substituents R₁, R₂, and R₃ are large hydrophobic groups. Similar results are also obtained for the spermidine series. Finally, differences in the interactions of the salts II and III with rA–rU₂ and rI–rC helices suggest that the latter helix is denser.     

Morphometric diffusing capacity and functional anatomy of the book lungs in the spider Tegenaria spp. (Agelenidae)

The presence of both book lungs and a tracheal system in many spiders raises the question of the functional significance of this double respiratory system. The present physiological and morphometric study of the house spider (Tegenaria spp.) reveals that the diffusing capacity (Dto₂) of the lungs alone suffices during rest and following exercise to meet measured rates of oxygen consumption (\documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm V}\limits^{\rm.} $\end{document}o₂) at driving pressures (ΔPto ₂) similar to those calculated for vertebrate lungs. During moulting ΔPto ₂ may rise to more than double the vertebrate values, implying the possible insufficiency of book lungs during this critical life phase. Resting \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm V}\limits^{\rm .} $\end{document}o₂ is greatest (92 mm³/h · g) during the early morning and lowest (66 mm³/h · g) near midday: during moulting \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm V}\limits^{\rm .} $\end{document}o₂ rises to 278.7 mm³/h · g. In spiders recovering from exercise \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm V}\limits^{\rm .} $\end{document}o₂ is consistently greater than during rest: neither value is significantly reduced by blockage of the tracheal stigmas. Regression calculations of morphometric values for a hypothetical 100-mg Tegenaria yield a total lung volume of 0.578 mm³, a pulmonary surface area of 69.8 mm², and a surface-to-volume ratio of 120.89 mm²/mm³. In spite of the similar thickness of the chitinous and hypodermal components of the air-hemolymph barrier (each ca. 0.2 μm in nonmoulting animals), the low permeability of chitin for oxygen makes this layer the greater barrier to diffusion. For a 100-mg specimen Dto₂ is 3.5 mm³/h · torr: similar to that of a turtle (Pseudemys) on a gram-body weight basis.     

Loss of ncm<Superscript>5</Superscript> and mcm<Superscript>5</Superscript> wobble uridine side chains results in an altered metabolic profile

Introduction

The Elongator complex, comprising six subunits (Elp1p-Elp6p), is required for formation of 5-carbamoylmethyl (ncm⁵) and 5-methoxycarbonylmethyl (mcm⁵) side chains on wobble uridines in 11 out of 42 tRNA species in Saccharomyces cerevisiae. Loss of these side chains reduces the efficiency of tRNA decoding during translation, resulting in pleiotropic phenotypes. Overexpression of hypomodified \( {\text {tRNA}_{{\rm s^{2} {\rm UUU}}}^{{\rm Lys}} , {\rm tRNA}_{{\rm s^{2} {\rm UUG}}}^{{\rm Gln }} \;{\rm and}\;{\rm tRNA}_{{\rm s^{2} {\rm UUC}}}^{{\rm Glu}}} \), which in wild-type strains are modified with mcm⁵s²U, partially suppress phenotypes of an elp3Δ strain.

Objectives

To identify metabolic alterations in an elp3Δ strain and elucidate whether these metabolic alterations are suppressed by overexpression of hypomodified \( {\text {tRNA}_{{\rm s^{2} {\rm UUU}}}^{{\rm Lys}} , {\rm tRNA}_{{\rm s^{2} {\rm UUG}}}^{{\rm Gln }} \;{\rm and}\;{\rm tRNA}_{{\rm s^{2} {\rm UUC}}}^{{\rm Glu}}} \).

Method

Metabolic profiles were obtained using untargeted GC-TOF-MS of a temperature-sensitive elp3Δ strain carrying either an empty low-copy vector, an empty high-copy vector, a low-copy vector harboring the wild-type ELP3 gene, or a high-copy vector overexpressing \( {\text {tRNA}_{{\rm s^{2} {\rm UUU}}}^{{\rm Lys}} , {\rm tRNA}_{{\rm s^{2} {\rm UUG}}}^{{\rm Gln }} \;{\rm and}\;{\rm tRNA}_{{\rm s^{2} {\rm UUC}}}^{{\rm Glu}}} \). The temperature sensitive elp3Δ strain derivatives were cultivated at permissive (30 °C) or semi-permissive (34 °C) growth conditions.

Results

Culturing an elp3Δ strain at 30 or 34 °C resulted in altered metabolism of 36 and 46 %, respectively, of all metabolites detected when compared to an elp3Δ strain carrying the wild-type ELP3 gene. Overexpression of hypomodified \( {\text {tRNA}_{{\rm s^{2} {\rm UUU}}}^{{\rm Lys}} , {\rm tRNA}_{{\rm s^{2} {\rm UUG}}}^{{\rm Gln }} \;{\rm and}\;{\rm tRNA}_{{\rm s^{2} {\rm UUC}}}^{{\rm Glu}}} \) suppressed a subset of the metabolic alterations observed in the elp3Δ strain.

Conclusion

Our results suggest that the presence of ncm⁵- and mcm⁵-side chains on wobble uridines in tRNA are important for metabolic homeostasis.

     

The effect of buffers and chelators on the reaction of luminol with Fenton's reagent near neutral pH

The chemiluminescence of the system luminol +Fe²⁺ + H₂O₂ was measured in aqueous buffer at pH 7.2. In veronal (5,5-diethybarbiturate) buffer, the luminescence is strongly quenched by ethanol and mannitol, but only weakly by t-butanol, benzoate and superoxide dismutase (SOD); complexing Fe²⁺ with 1,10-phenanthroline or 2,2′-dipyridyl causes a decrease of light production that can be partially obviated by the simultaneous addition of SOD. In phosphate buffer, the luminescence is higher than in veronal and it is efficiently quenched by all four OH · quenchers and by SOD. In Tris buffer, no light production is observed as long as the Fe²⁺ is not complexed. When Fe²⁺ is complexed by pyrophosphate or phytate, there is a strong chemiluminescence in all three buffers, which is quenched by all four OH · quenchers and by SOD. When Fe²⁺ is complexed by EDTA or DTPA, very little luminescence is observed. The luminol analogue phthalhydrazide, which was suggested by Merényi and Lind as a reliable OH · detector, can replace luminol only in phosphate buffer, and thus turns out to be very specific indeed for free OH ·.     

The humus layer determines SO<Subscript>4</Subscript><Superscript>2−</Superscript> isotope values in the mineral soil

Biocycling of sulfur (S) has been proposed to play an important role in the recovery of ecosystems following anthropogenic S deposition. Here, we investigated the importance of the humus layer in the biocycling of S in three forested catchments in the Gårdsjön area of southwestern Sweden with differing S inputs and S isotope signature values. These experimental sites consisted of two reference catchments and the Gårdsjön roof experiment catchment (G1), where anthropogenic deposition was intercepted from 1991 until May 2002 by a roof placed over the entire catchment area. Under the roof, controlled levels of deposition were applied, using a sprinkler system, and the only form of S added was marine SO₄²⁻ with a δ of +19.5‰.We installed ion exchange resin bags at the interface between the humus layer and mineral soil at each of the catchments to collect SO₄²⁻ passing through the humus. The resin bags were installed on four occasions, in 1999 and 2000, covering two summer and two winter periods. The ions collected by each bag during these sampling periods were then eluted and their δ values and SO₄²⁻ concentrations determined. The most striking result is that the average δ value in the resin bags was more than 12‰ lower compared to that of the sprinkler water in the G1 roof catchment. There was no increasing trend in the isotope value in the resin bag SO₄²⁻ despite that the roof treatment has been on-going for almost 10 years; the average value for all resin bags was +7.1‰. The highest δ values found in the G1 roof catchment were between +11‰ and +12‰. However, these values were all obtained from resin bags installed at a single sampling location. Throughfall and resin bag δ values were more similar in the two reference catchments: about +7.5‰ in both cases. There was, however, an increase in resin bag δ values during the first winter period, from about +7‰ to +9‰. The resin bag δ value was linearly and positively related (r² = 0.26, p < 0.001) to the amount of SO₄²⁻ extracted from the resin bags, if relatively high amounts (>50 mmol m⁻²) were excluded. High amounts of resin bag SO₄²⁻ seemed to be related to groundwater inputs, as indicated by the δ value. Our results suggest that rapid immobilization of SO₄²⁻ into a large organic S pool may alter the S isotope value and affect the δ values measured in the mineral soil and runoff.     

Sequence and age of eruption of deciduous dentition in the baboon (Papio sp)

A detailed eruption sequence and associated age of eruption for deciduous dentition in baboons (Papio sp) are presented in this paper. The sequence was determined by evaluation and comparison of the number and kinds of teeth present in nine age cohorts comprising the study sample of 88 males and 87 females who ranged in age from birth to 763 days. Eruption was assessed visually as present or absent. Several statistical methods used to derive the ages associated with the eruption sequence are described. The basic eruption sequence in the sample population is: i¹ i₁, i₂, i², \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm c}\limits_{\rm -} {\rm,}\mathop {\rm c}\limits^{\rm -} $\end{document} m¹ (m², m₂), M₁, M¹. Both sexes show the same pattern, with the exception of the second deciduous molar, where males show a sequence of m₂, m₂, while females show the opposite. Posterior dentition shows the greatest gender-specific variation in average age of eruption.     

Photosynthesis in Pineapple (Ananas comosus comosus [L.] Merr) Measured Using PAM (Pulse Amplitude Modulation) Fluorometry

PAM (Pulse Amplitude Modulation) fluorometer techniques directly measure the light reactions of photosynthesis that are otherwise difficult to estimate in CAM (Crassulacean Acid metabolism) plants such as pineapple (Ananas comosus comosus cv. Phuket). PAM machines calculate photosynthesis as the Electron Transport Rate (ETR) through PSII (4 electrons per O₂ produced) as mol m^?2 s^?1. P vs. E curves fitted the waiting-in-line function (an equation of the form $ {\hbox{ETR}} = \left( {{\hbox{ET}}{{\hbox{R}}_{{ \max }}} \times {\hbox{E}}/{{\hbox{E}}_{\rm{opt}}}} \right).{{\hbox{e}}^{{1} - {\rm{E}}/{\rm{Eopt}}}} $ ) allowing half-saturating and optimal irradiances (E_opt) to be estimated. Effective Quantum Yield (Y_max), Electron Transport Rate (ETR_max) and the Non-Photochemical Quenching parameter, NPQ_max all vary on a diurnal cycle but the parameter qN_max does not show a systematic variation over a diurnal period. Phuket pineapple is a “sun plant” with Optimum Irradiance (E_opt) from 755 to 1,130 μmol m^?2 s^?1 (400–700 nm) PAR but photosynthetic capacity is very low in the late afternoon even though light conditions are favourable for rapid photosynthesis. Total CO₂ fixed nocturnally as C4-dicarboxylic acids by leaves of the Phuket pineapple was only ≈0.14 gC m^?2 d^?1 (0.012 mol C m^?2 d^?1). Titratable acid of leaves was depleted about 3 pm (15:00) and shows a classical CAM diurnal cycle. The Phuket pineapple variety only stored enough CO₂ as C4 acids to account for only about 2.5% of photosynthesis (P_g) estimated using the PAM machine (≈5.6 gC m^?2 d^?1). Phuket pineapples are classifiable as CAM-Cycling plants but nocturnal fixation of CO₂ is so low compared to the more familiar Smooth Cayenne variety that it probably recycles only a small proportion of the respiratory CO₂ produced in leaves at night and so even CAM-cycling is only of minor importance to the carbon economy of the plant. Unlike the Smooth Cayenne pineapple variety, which fixes large amounts of CO₂ nocturnally, the Phuket pineapple is for practical purposes a C3 plant.     

Toxicity of oxygen from naturally occurring redox-active pro-oxidants

The survival of all aerobic life forms requires the ground-state of molecular oxygen, O₂. However, the activation of O₂ to reactive oxygen species (ROS) is responsible for universal toxicity. ROS are responsible in deleterious intracellular reactions associated with oxidative stress including membrane lipid peroxidation, and the oxidation of proteins and DNA. Redox-active allelochemicals such as quinones and phenolic compounds are involved in activating O₂ to its deleterious forms including superoxide anion free radical, $ {\rm O}_{\rm 2} ^{ \cdot - } $, hydrogen peroxide, H₂O₂, and hydroxyl radical, $ \cdot {\rm OH} $. Molecular oxygen is also activated in biologically relevant photosensitizing reactions to the singlet form, ¹O₂. The insect lifestyle exposes them to a broad diversity of pro-oxidant allelochemicals and, like mammalian species, they have developed an elaborate antioxidant system comprised of chemical antioxidants and a bank of antioxidant enzymes. We have found that an insect's antioxidant adaptation to a particular food correlates well with its risk of exposure to potential pro-oxidants. © 1995 Wiley-Liss, Inc.     

Measurement of eight scalar and dipolar couplings for methine–methylene pairs in proteins and nucleic acids

A new 3D, spin-state-selective coherence transfer NMR experiment is described that yields accurate measurements for eight scalar or dipolar couplings within a spin system composed of a methylene adjacent to a methine group. Implementations of the experiment have been optimized for proteins and for nucleic acids. The experiments are demonstrated for C–C moieties of the third IgG-binding domain from Streptococcal Protein G (GB3) and for C –C groups in a 24-nt RNA oligomer. Chemical shifts of C, C and H (respectively C , C and H ) are dispersed in the three orthogonal dimensions, and the absence of heteronuclear decoupling leads to distinct and well-resolved E.COSY multiplet patterns. In an isotropic sample, the E.COSY displacements correspond to ¹J_CH, ²J_CH2+²J_CH3, ²J_CH, ¹J_CH2+¹J_CH3, ¹J_CH2–²J_H2H3, ¹J_CH3–²J_H2H3, ³J_HH2 and ³J_HH3 for proteins, and ¹J , ²J J , ²J , ¹J +¹J , ¹J J , ¹J J , ³J and ³J in nucleic acids. The experiment, based on relaxation-optimized spectroscopy, yields best results when applied to residues where the methine–methylene group corresponds to a reasonably isolated spin system, as applies for C, F, Y, W, D, N and H residues in proteins, or the C –C groups in nucleic acids. Splittings can be measured under either isotropic or weakly aligned conditions, yielding valuable structural information both through the ³J couplings and the one-, two- and three-bond dipolar interactions. Dipolar couplings for 10 out of 13 sidechains in GB3 are found to be in excellent agreement with its X-ray structure, whereas one residue adopts a different backbone geometry, and two residues are subject to extensive ₁ rotamer averaging. The abundance of dipolar couplings can also yield stereospecific assignments of the non-equivalent methylene protons. For the RNA oligomer, dipolar data yielded stereospecific assignments for six out of the eight C H₂ groups in the loop region of the oligomer, in all cases confirmed by ¹J ^{1} $$" align="middle" border="0"> J , and H resonating downfield of H .Supplementary material to this paper is available in electronic form at http://dx.doi.org/10.1007/s10858-005-0175-z.     

Adiabatic transformability hypothesis of human locomotion

It is hypothesized that metabolic and mechanical changes in human locomotion associated with changes in speed v are constrained by two attractive strategies: $Q_{{\text{metab}}} = 1{\text{ and }}\Delta Q_{{\text{metab}}} /\Delta v = {\text{a}}$ positive definite constant. $Q_{{\text{metab}}} = \Delta {\rm E}_{\text{k}} {\text{s}}^{{\text{ - 1}}} /{\text{ml O}}_{\text{2}} {\text{s}}^{{\text{ - 1}}} $ where ΔEs^?1 is the summed increments and decrements per unit time in the translational and rotational kinetic energies of the body's segments and ml O₂s^?1 is the rate at which chemical energy is dissipated. The expected constancy of ΔQ _metab/Δv _metab was derived from an extension of Ehrenfest's adiabatic hypothesis by which transformations (increases, decreases) in locomotion v can be considered as adiabatic, even though the biological conditions are nonconservative and non-rate-limited. The expected significance of Q _metab=1 was derived from stability considerations of the symmetry per stride of stored and dissipated energy. An experimental evaluation was provided by collecting metabolic and mechanical measures on walking (10 subjects) and running (9 subjects) at progressively greater treadmill speeds but within the aerobic limit. Results revealed that walking was restricted to ometab ? 1 with a nonlinear trajectory in v×Q _metab coordinates shaped by Q _metab=1 (primarily) and the constancy of ΔQ _metab/Δv. Running satisfied Q _metab > 1, with a linear trajectory in v×Q _metab coordinates conforming to ΔQ _metab/_Δv=a constant, with the constant predicted from invariants in the mechanical space v×ΔE _ks^?1. Results also suggested that the metabolic costs of running might be predictable from measures made in the v×ΔE _ks^?1 space.     

Report on non-temperature related variations in CO<Subscript>2</Subscript> efflux rates from young tree stems in the dormant season

Respiration rates are reported to increase exponentially with temperature. Respiration rates of woody tissues are commonly measured as CO₂ efflux rates () from that tissue. However, this paper describes clear variations in stem that were not related to temperature for the case of a young beech (Fagus sylvatica L.) and oak (Quercus robur L.) tree during the dormant season. The CO₂ concentration ([CO₂]) in the xylem of the beech tree showed similar temperature-independent variations. The trees were grown in a growth chamber in which radiation patterns and temperature were kept constant. was measured with an IRGA connected to cuvettes surrounding a stem segment. Xylem [CO₂] was measured in situ using a CO₂ microelectrode. Depressions in and [CO₂] occurred during the light period, despite equal temperatures in the light and dark period. Explanations found in literature for discrepancies in the exponential relationship between temperature and are the influence of (1) sap flow or (2) decreased cell water content. However, (1) the variations were observed in the dormant season, when no sap flow was observed yet, and (2) reduced cell water content was not likely to be apparent as differences in stem transpiration rates between the dark and light period were not significant. Hence, previously formulated theories failed to explain our results. This work therefore provides a new ground for discussion on other possible causes of daytime depressions in . One might be the refixation of respired CO₂ by corticular photosynthesis in the stem parts adjacent to the stem segment enclosed by the cuvette.     

A set of HA-detected experiments for measuring scalar and residual dipolar couplings

A new set of HCACO based three-dimensional NMR experiments for measuring residual dipolar couplings in proteins is presented. Using spin-state selection and editing in three dimensions, the experiments allow accurate measurement of intraresidual , and scalar and residual dipolar couplings of ¹⁵N/¹³C labeled proteins in D₂O and dilute liquid crystals with minimal spectral crowding. The presented experiments are especially suitable for small or medium sized proline-rich proteins, or proteins that require high pH solvent conditions, making ¹H^N detected experiments unattractive. In addition, the tetrahedral coordination of C is superior to the planar peptide bond for determination of local alignments in partially structured polypeptides. For the efficient use of spectrometer time and to avoid complications arising from the varying magnitude of the alignment tensor during relatively long experiments, the and couplings can also be measured simultaneously in an E.COSY like manner with high accuracy. The pulse sequences are balanced for cross-correlation effects and minimized for relaxation losses. The pulse sequences are tested with a sample of ¹⁵N/¹³C human ubiquitin. We find internuclear vector directions determined from the dipolar couplings to have an excellent correlation with those of ubiquitins refined solution structure.     

Formate binding near the redox-active TyrosineD in Photosystem II: consequences on the properties of TyrD

Formate and phosphate affect substantially the rate of tyrosine D (Tyr_D) oxidation and the stability of the radical Tyr in Photosystem II [Hienerwadel R, Boussac A, Breton J and Berthomieu C (1996) Biochemistry 35: 15447–15460]. This observation prompted us to analyze the influence of formate and phosphate on the environment of Tyr_D using FTIR spectroscopy. The ν (CO) IR mode of Tyr at 1503 cm⁻¹ remains unchanged whatever the buffer used at pH 6 and whether formate is present or not in the sample. Similarly, the main IR mode of reduced Tyr_D remains at ≈1250 cm⁻¹ in all tested conditions. We thus conclude that formate does not modify the hydrogen-bonded interactions of Tyr_D and Tyr with neighbouring D2His189 and D2Gln164. In the Tyr_D-state, an IR mode of formate significantly different from that observed in solution, is detected using ¹³C-formate, showing that formate forms a strong electrostatic interaction within PS II. The presence of formate affects also IR bands that may be assigned to an arginine side chain. Upon Tyr formation, formate does not protonate but its binding interaction weakens. A proton uptake by Mes or phosphate buffer is detected, which is not observed when BisTris is used as a buffer. In these latter conditions, IR bands characteristic of the protonation of a carboxylate group of the protein are detected instead. The present IR data and the recent structural model of the Tyr_D environment proposed by Ferreira KN, Iverson TM, Maghlaoui K, Barber J and Iwata S [(2004) Science 303: 1831–1838], suggest that the proton released upon Tyr formation is shared within a hydrogen bonding network including D2Arg294, and CP47Glu364 and that perturbation of this network by formate – possibly binding near D2Arg294 – substantially affects the properties of Tyr_D.     

Die Erzeugung von Chemilumineszenzen wÄ\riger TlI-Salzlösungen durch Röntgenstrahlen

Zusammenfassung Die Lumineszenz wÄ\riger Tl⁺-Lösungen unter Einwirkung von 8,5-keV_eff-Röntgenstrahlen wird untersucht, und zwar 1. in neutraler Lösung, 2. in saurer Lösung, 3. in alkalischer Lösung, 4. bei Zusatz von NaCl.Bei Änderung des pH-Wertes nimmt die Lichtausbeute sowohl auf der sauren Scite (beipH=3,4) als auch auf der alkalischen Scite (beipH=12) ein Maximum an; sie erreicht dort den 2- bis 3fachen Wert der Ausbeute in neutraler Lösung. Andererseits ist bei Zusatz von 1 M NaCl in neutraler Lösung die Ausbeute fünfmal höher als bei Abwesenheit von NaCl; die Maxima im sauren und alkalischen Bereich verschwinden jedoch bei 1 M NaCl vollkommen, und man erhÄlt bei gro\en und bei kleinen pH-Werten lediglich eine Löschung der Lumineszenz.Die Lumineszenz kommt durch Bildung von Tl^+* bei der Anlagerung von e_aq an Tl⁺⁺ zustande. Tl⁺⁺ entsteht jedoch nicht unmittelbar gemÄ\ Tl⁺ + OH Tl⁺⁺ + OH^–, sondern zunÄchst bildet sich Tl⁺OH, und dieses dissoziiert erst nach einer Verzögerung von > 10^–7 sec in Tl⁺⁺ und OH^–. Die Sensibilisierung durch H⁺ wird durch die Reaktion Tl⁺OH + H⁺ Tl⁺⁺ + H₂O und die durch OH^– durch Tl⁺OH + OH^– Tl⁺⁺O^– + H₂O erklÄrt. Die Reaktion der Komplexe Tl⁺Cl^– und Tl⁺Cl₂ ^– mit OH erfolgt offenbar ohne zeitliche Verzögerung, d. h. Tl⁺⁺Cl^–– bzw. Tl⁺⁺Cl₂ ^–– wird dabei innerhalb einer Zeit 10^–7 sec gebildet.

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Chemiluminescences of aqueous Tl^I solutions produced by irradiation with 8.5-keV-X-rays

Summary The luminescence of aqueous solutions of Tl⁺ during irradiation with 8.5-keV-X-rays has been investigated. In particular we have studied the Tl⁺-luminescence in 1. neutral solutions, 2. acid solutions, 3. alkaline solutions, 4. solutions containing various concentrations of NaCl.Varying thepH the luminescence yield passes a maximum on the acid side as well as on the alkaline side (atpH=3.4 respectivelypH=12). Compared with neutral solutions the luminescence yield is increased by a factor 2 to 3 at these pH-values.By adding 1 M NaCl to neutral Tl⁺-solutions the luminescence yield is enhanced by a factor five, however in dependence ofpH no further increase has been observed, but only quenching of the luminescence at low and highpH.The luminescence origins from the formation of Tl^+* by the reaction of e_aq with Tl⁺⁺ formed by oxidation of Tl⁺ by OH. However, Tl⁺⁺ is not formed directly by Tl⁺+OH Tl⁺⁺ + OH^– but via dissociation of the intermediate Tl+OH after a delay > 10^–7 sec.We explain the increase of the luminescence yield in acid solutions by the reaction Tl⁺OH + H⁺ Tl⁺⁺ + H₂O and in alkaline solutions by the reaction Tl⁺OH + OH^– Tl⁺⁺O^– + H₂O. In alkaline solutions the luminescence spectrum shifts to longer wavelengths; we conclude, that this spectrum is attributed to Tl^+*O^–. Evidently the reaction of Tl+Cl^– and Tl+Cl₂ ^–– with OH leads to formation of Tl⁺⁺Cl^– and Tl⁺⁺Cl₂ ^–– without any efficient delay.

     

Streptomyces daliensis sp. nov. from soil

A novel actinomycete strain YIM 31724^T was isolated from a soil sample collected from Dali, Yunnan Province, People’s Republic of China. The strain is characterized by white to yellow white aerial mycelia, spiral spore chains and smooth spore surface. The cell wall of strain YIM 31724^T contained LL-diaminopimelic acid (A₂pm) and traces of meso-A₂pm. Whole-cell hydrolysates contained mainly glucose and small amounts of galactose and xylose. The menaquinones were MK-9(H₆) (31%) and MK-9(H₈) (69%). Phosphatidylethanolamine was the diagnostic phospholipid. The DNA G+C content of strain YIM 31724^T was 67.2 mol%. Phylogenetic analysis indicated that the strain belongs to the genus Streptomyces, with highest similarity to Streptomyces rimosus subsp. rimosus JCM 4667^T (rRNA gene sequence similarity value of 98.9%) and Streptomyces erumpens DSM 40941^T (rRNA gene sequence similarity value of 98.7%). Based on its phenotypic and genotypic characteristics, including low DNA–DNA hybridization results, strain is proposed as the type strain of a novel species, Streptomyces daliensis sp. nov.     

The regulation of K+ influx in excised barley roots

The electrochemical potential differences for potassium, between excised barley (Hordeum vulgare L.) roots and external media containing 0.05 mM KCl+0.5 mM CaSO₄, were determined over a 4-h period during which initially low-K⁺ roots accumulated K⁺ by pretreatment in 50 mM KCl plus 0.5 mM CaCl₂. This pretreatment resulted in increased internal [K⁺], decreased K⁺ influx (as measured from 0.05 mM KCl+0.5 mM CaSO₄) and decreased values of . These observations indicate that the decline of K⁺ influx associated with increased internal K⁺ concentration cannot be accounted for by passive adjustment to the electrochemical gradient for this ion.     

Magnetic resonance and kinetic studies of the mechanism of membrane-bound sodium and potassium ION-activated adenosine triphosphatase

EPR and water proton relaxation rate (1/T₁) studies of partially (40%) and “fully” (90%) purified preparations of membrane-bound (Na⁺+K⁺) activated ATPase from sheep kidney indicate one tight binding site for Mn²⁺ per enzyme dimer, with a dissociation constant (K_D = 0.88 μM) in agreement with the kinetically determined activator constant, identifying this Mn²⁺-binding site as the active site of the ATPase. Competition studies indicate that Mg²⁺ binds at this site with a dissociation constant of 1 mM in agreement with its activator constant. Inorganic phosphate and methylphosphonate bind to the enzyme-Mn²⁺ complex with similar high affinities and decrease l/T₁ of water protons due t o a decrease from four to three in the number of rapidly exchanging water protons in the coordination sphere of enzyme-bound Mn²⁺. The relative effectiveness of Na⁺ and K⁺ in facilitating ternary complex formation with HPO and CH₃PO as a function of pH indicates that Na⁺ induces the phosphate monoanion t o interact with enzyme-bound Mn²⁺, while K⁺ causes the phosphate dianion to interact with the enzyme-bound Mn²⁺. Thus protonation of an enzyme-bound phosphoryl group would convert a K⁺-binding site to a Na⁺-binding site. Dissociation constants for K⁺ and Na⁺, estimated from NMR titrations, agreed with kinetically determined activator constants of these ions consistent with binding t o the active site. Parallel ³²P_i-binding studies show negligible formation (< 7%) of a covalent E–P complex under these conditions, indicating that the NMR method has detected an additional noncovalent intermediate in ion transport. Ouabain, which increases the extent of phosphorylation of the enzyme to 24% at pH 7.5 and t o 106% at pH 6.1, produced further decreases in l/T 1 of water protons. Preliminary ³¹P-relaxation studies of CH₃PO in the presence of ATPase and Mn²⁺ yield an Mn to P distance (6.9 ± 0.5 Å) suggesting a second sphere enzyme-Mn-ligand-CH₃PO complex. Previous kinetic studies have shown that T1⁺ substitutes for K⁺ in the activation of the enzyme but competes with Na⁺ at higher levels. From the paramagnetic effect of Mn²⁺ at the active site on the enzyme on I/T₁ of ²⁰⁵T1 bound at the Na⁺ site, a Mn²⁺ to T1⁺ distance of 4.0 ± 0.1 Å is calculated, suggesting the sharing of a common ligand atom by Mn²⁺ and T1⁺ on the ATPase. Addition of P. increases this distance to 5.4 Å consistent with the insertion of P between Mn²⁺ and T1⁺. These results are consistent with a mechanism for the \documentclass{article}\pagestyle{empty}\begin{document}$ (\mathop {\rm N}\limits^{\rm i} {\rm a}^{\rm + } {\rm + K}^ +) $\end{document}-ATPase and for ion transport in which the ionization state of Pi at a single enzyme active site controls the binding and transport of Na⁺ and K⁺, and indicate that the transport site for monovalent cations is very near the catalytic site of the ATTase. Our mechanism also accounts for the order of magnitude weaker binding of Na⁺ compared to K⁺.     

The intramolecular δ<Superscript>15</Superscript>N of lysine responds to respiratory status in <Emphasis Type="Italic">Paracoccus denitrificans</Emphasis>

Summary. Presented here is the first experimental evidence that natural, intramolecular, isotope ratios are sensitive to physiological status, based on observations of intramolecular δ¹⁵N of lysine in the mitochondrial mimic Paracoccus denitrificans. Paracoccus denitrificans, a versatile, gram-negative bacterium, was grown either aerobically or anaerobically on isotopically-characterized ammonium as sole cell-nitrogen source. Nitrogen isotope composition of the biomass with respect to source ammonium was = −6.2 ± 1.2‰ for whole cells under aerobic respiration, whereas cells grown anaerobically produced no net fractionation ( = −0.3 ± 0.23‰). Fractionation of ¹⁵N between protein nitrogen and total cell nitrogen increased during anaerobic respiration and suggests that residual nitrogen-containing compounds in bacterial cell membranes are isotopically lighter under anaerobic respiration. In aerobic cells, the lysine intramolecular difference between peptide and sidechain nitrogen is negligible, but in anaerobic cells was a remarkable Δ¹⁵N_{p − s} = δ¹⁵N_peptide − δ¹⁵N_sidechain = +11.0‰, driven predominantly by enrichment at the peptide N. Consideration of known lysine pathways suggests this to be likely due to enhanced synthesis of peptidoglycans in the anaerobic state. These data indicate that distinct pathway branching ratios associated with microbial respiration can be detected by natural intramolecular Δδ¹⁵N measurements, and are the first in vivo observations of position-specific measurements of nitrogen isotope fractionation.     

Topography of nucleic acid helices in solutions. II. Structure of the double-stranded rA-rU, rI-rC, acid rA, and the triple-stranded rA-rU2 and rA-rI2 helices

Information concerning the structures of rA–rU, rA–rU₂ rI–rC, rA–rI₂, and acid rA helices in solutions is reported. Through the use of diquaternary ammonium salts of the general structure, \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm R}_1 {\rm R}_2 {\rm R}_3 \mathop {\rm N}\limits^ + ({\rm CH}_2 )n\mathop {\rm N}\limits^ + {\rm R}_1 {\rm R}_2 {\rm R}_3 \cdot 2{\rm Br}^ - $\end{document} (I), it is shown that (1) the distances between adjacent negatively charged oxygen atoms on the helix increases in the following order rA–rI₂ < rI–rC < rA–rU ? rA–rU₂; (2) the density of the helices increases in the order. rA–rI₂ < rA–rU < rA–rU₂ < rI–rC; (3) there is a large hydrophobia site in rA–rI₂ and possibly also in rA–rU, rA–rU₂, and rI–rC helices; (4) the results of the interactions between the salts of type I and the helices may be formulated in semi-quantitative terms by the use of two parameters, α, and β which are shown to be related to the charge separation and the density of the helices, respectively; (5) the studies in solutions compare favorably with the x-ray studies on the fibers; and (6) the acid rA helix differs significantly from the other helices by the fact that the electrostatic interstrand interactions between the negatively charged oxygen atom of a phosphate group and the positively charged 10-amino group of adenine contribute significantly to the stabilization of the helix, and thus it is found that the presence of the salts, I, leads to a significant destabilization of the acid rA helix.