The mechanism of deuterium oxide-induced protein degradation in Lemna minor

Transfer of Lemna minor fronds to culture medium containing 50% (v/v) deuterium oxide induces a large increase in the rate of protein breakdown, which is not due to an increase in the activity of acidic or neutral proteolytic enzymes or peptidases. Biochemical and ultrastructural evidence indicates that deuterium oxide affects the properties of certain membranes, particularly the tonoplast, and allows vacuolar proteolytic enzymes to pass into the cytoplasm and cause the increased protein breakdown.Abbreviations BAPA benzylarginine-p-nitroanilide - LPA leucine-p-nitroanilide - TCA trichloroacetic acid     

Passive cable properties of hippocampal CA3 pyramidal neurons

The passive electrical cable properties of CA3 pyramidal neurons from guinea pig hippocampal slices were investigated by applying current steps and recording the voltage transients from 25 CA3 neurons, using a single intracellular microelectrode and a 3-kHz time-share system. Two independent methods were used for estimating the equivalent electrotonic length of the dendrites, L, and the dendritic to somatic conductance ratio, . The first method is similar to that used by Gorman and Mirolli (1972) and gave an average L of 0.96; the average was 2.44. The second method is derived here for the first time and assumes a finite-length cable with lumped soma. It is an exact solution for L and , using the slopes and intercepts of the first two peeled exponentials. The average L was 0.94; the average was 1.51. The results, using both methods, are in close agreement. The average membrane time constant for all 25 CA3 neurons was 23.6 ms, suggesting a large (23,600 cm²) average membrane resistivity. It is concluded that CA3 neurons are electronically short.This work was supported by Grants NS 11535 and NS 15772 from the National Institute of Neurological and Communicative Disorders and Stroke, National Institutes of Health, U.S. Public Health Service.     

Purification and properties of O-acetyl-L-serine sulphydrylase from wheat leaves

O-Acetyl-L-serine sulphydrylase (OASS), the enzyme which produces L-cysteine from O-acetyl-L-serine (OAS) and sulphide, is     

An unusual allantoinase from Dolichos biflorus

An unusual allantoinase from Dolichos biflorus has been purified 62-fold. The purified enzyme has an unusual pH activity profile with a shoulder at pH 4 and a peak at pH 7.5. This is due to a single enzyme which does not need metal ions for activation. In the fully reduced state the enzyme exhibits a single sharp peak at 7.5; when it is not in the sulfhydryl form (in the fully oxidized SS form?) the enzyme shows a single pH optimum at pH 4. K_m values for (±)-allantoin were 5.5 mM at pH 4 and 1.43 mM at pH 7.5. Allantoinase activity has been demonstrated in the resting seed, and increased linearly with time during the first 5 days of seedling growth.     

An aminopeptidase from Agave americana thermodynamic studies

Purified Agave aminopeptidase was characterized with respect to the thermodynamic properties of the reaction catalysed by the enzyme. Kinetic studies were conducted at different temperatures ranging from 7.8 to 45°. The energy of activation, E_a, as well as the constants ΔH, ΔF and ΔS were calculated for both the formation of the enzyme-substrate complex, and the dissociation of the enzyme-product complex. Kinetic studies in buffers with varying dielectric constants enabled the determination of the electrostatic as well as the non-electrostatic components of ΔS. These results fit well into the overall kinetic picture of this enzyme-catalysed reaction as reported previously.     

Purification and properties of mannitol dehydrogenase from Agaricus bisporus sporocarps

Mannitol dehydrogenase (mannitol: NADP⁺ 2-oxidoreductase: EC 1.1.1.138) was isolated from Agaricus bisporus by fractionation with protamine sulphate and (NH₄)₂SO₄, followed by chromatography on DEAE-Sephadex, then by affinity and gel chromatography. The products of enzyme reaction were identified by GLC and TLC. K_m, optimum pH, MW and pI of the enzyme as well as the influence of temperature, ions and inhibitors on enzymic activity were determined. In the sugar reducing reaction, the enzyme was specific for fructose but, in the reverse direction, some structurally related polyols could substitute for mannitol. The enzyme was very sensitive to alterations in the NADP⁺/NADPH ratio. The results are discussed in relation to the possible role of mannitol dehydrogenase in fungal metabolism.     

Cinnamic acid 4-hydroxylase from gherkin tissues

Homogenates from 4-day-old gherkin cotyledons and hypocotyls fractionated by sucrose density gradient centrifugation contain cinnamic acid 4-hydroxylase activity, the activity being highest in the endoplasmic reticulum fractions. These fractions also contain very low concentrations of cytochrome P₄₅₀. Hydroxylase activity is dependent on NADPH and on molecular oxygen, is optimal at pH 7.5 and is inhibited by carbon monoxide. The enzyme is very sensitive to inhibition by 2-mercaptoethanol, but it is not inhibited by the product, p-coumaric acid. Further, its responses to various potential inhibitors are fairly typical of mixed function oxidases from other sources.     

Alginate as immobilization material: III. Diffusional properties

The rate of diffusion of serum albumin (MW 6.9 x 10(4) D) out of beads of calcium alginate gels depends upon the concentration and uronic acid composition of the alginate (ManA/GulA ratio), the conditions under which the beads are produced, the pH, and the temperature. The diffusion coefficient decreases with increasing alginate concentration, and (ManA/GulA) ratio and with decreasing pH. Diffusion out of the beads, in which the alginate is uniformly distributed (homogeneous gel), is faster than out of the beads in which the alginate is concentrated at the surface (inhomogeneous gel). The temperature dependence of the diffusion coefficient follows the Arrhenius law, with an activation energy of approximately 23 kJ x mol(-1).     

Are some plant life forms more effective than others in screening out ultraviolet-B radiation?

Summary The unprecedented rate of depletion of the stratospheric ozone layer will likely lead to appreciable increases in the amount of ultraviolet-B radiation (UV-B, 280–320 nm) reaching the earth's surface. In plants, photosynthetic reactions and nucleic acids in the mesophyll of leaves are deleteriously affected by UV-B. We used a fiber-optic microprobe to make direct measurements of the amount of UV-B reaching these potential targets in the mesophyll of intact foliage. A comparison of foliage from a diverse group of Rocky Mountain plants enabled us to assess whether the foliage of some plant life forms appeared more effective at screening UV-B radiation. The leaf epidermis of herbaceous dicots was particularly ineffective at attenuating UV-B; epidermal transmittance ranged from 18–41% and UV-B reached 40–145 m into the mesophyll or photosynthetic tissue. In contrast to herbaceous dicots, the epidermis of 1-year old conifer needles attenuated essentially all incident UV-B and virtually none of this radiation reached the mesophyll. Although the epidermal layer was appreciably thinner in older needles (7 y) at high elevations (Krumholtz), essentially all incident UV-B was attenuated by the epidermis in these needles. The same epidermal screening effectiveness was observed after removal of epicuticular waxes with chloroform. Leaves of woody dicots and grasses appeared intermediate between herbaceous dicots and conifers in their UV-B screening abilities with 3–12% of the incident UV-B reaching the mesophyll. These large differences in UV-B screening effectiveness suggest that certain plant life forms may be more predisposed than others to meet the challenge of higher UV-B levels resulting from stratospheric ozone depletion.     

Chirality of reverse micelles

Four chiral analogues of the surfactant Aerosol-OT (AOT) have been synthesized and characterized. All of them form reverse micelles in apolar solvents in the w₀ range 0–30 (w₀ = [water]/[tenside]). Reverse micellar solutions have been investigated by UV absorption and circular dichroism spectroscopies with the aim of clarifying whether the formation of the macromolecular micellar structure induces the appearance of new chromophoric bands or perturbs the existing ones. Methanolic solutions of the surfactants, in which no micellar aggregates are formed, were taken as references. One of the products 1(S),1′(S)-dimethylbisheptylsulphosuccinate sodium salt (MH-AOT) was capable of forming reverse micelles of relatively high water content (w₀ up to 40) and this process was accompanied by a specific increase in the intensity of the circular dichroism band associated with the ester absorbance of the molecule. As no concomitant changes were seen in the UV absorbance spectrum, it was concluded that this observation reflected conformational events occurring within the surfactant rather than chromophoric perturbation. These results are qualitatively similar to those found recently for lecithin reverse micelles which, however, form gels at sufficiently high water contents. The chiroptical properties of these supramolecular aggregates are compared with those of covalent macromolecular systems such as polypeptides.