NMR characterization of three forms of ferredoxin from Desulphovibrio gigas, a sulphate reducer

A NMR and magnetic susceptibility study of the oxidized and reduced states of three different oligomers (forms) of a [4Fe-4S] ferredoxin protein from Desulphovibrio gigas, FdI, FdI′, and FdII was carried out. FdI and FdI′ are different trimers and FdII a tetramer of the same basic subunit. A probable assignment of the contact shifted resonances is indicated. Since the temperature dependences of the contact shifted resonances associated with each [4Fe-4S] are not all similar a delocalized model for the spin densities on the 4Fe does not apply. The exchange rate between oxidized and reduced states is slow on the NMR time scale. The three oligomers are not magnetically equivalent. Using the “three state hypothesis” terminology it is shown that FdI_ox is predominantly in the C^2? state and changes upon reduction into the C^3? state, while FdII_ox is in the C^? state and changes into the C^2? state. FdI′ does not easily fit into this classification. This study shows a similarity of magnetic behaviour between FdI and bacterial ferredoxins (e.g. Bacillus polymyxa) and between FdII and HiPIP from Chromatium sp.. The influence of the quaternary structure on the stabilization of the different oxidation states of ferredoxins as well as on their redox potentials is discussed.     

Variations in litter size and reproductive effort within and between some populations of Lacerta vivipara

We studied the relationships between litter size, litter weight, newborn weight, relative clutch mass and the female snout-vent length in some Lacerta vivipara populations over a period of three years.
Litter size and litter weight were positively correlated with female snout-vent length in all the populations for all the years, as in most other lizard species. Relative clutch mass generally increased with female size, though correlations appear not to be very tight.
Considering the two best studied populations suggests that montane females invest less in reproduction than lowland ones.
The main reproductive traits of the species appeared highly variable between as well as within the different populations hitherto studied.
We argue that current theory about lizard reproductive strategy requires, first to work out a good estimate of reproductive effort, and second to get more information about the relations between the species and their environmental, biotic and abiotic conditions.     

NMR redox studies of Desulfovibrio vulgaris Cytochrome c3. Electron transfer mechanisms

The 300-MHz proton NMR spectra of the tetrahaem cytochrome c3 from Desulfovibrio vulgaris were examined while varying the pH and the redox potential. The analysis of the complete NMR reoxidation pattern was done taking into account all the 16 redox states that can be present in the redox titration of a tetra-redox-center molecule. A network of saturation transfer experiments performed at different oxidation stages, between the fully reduced and the fully oxidized states, allowed the observation of different resonances for some of the haem methyl groups. In the present experimental conditions, some of the haems show a fast intramolecular electron exchange rate, but the intermolecular electron exchange is always slow. In intermediate reoxidation stages, large shifts of the resonances of some haem methyl groups were observed upon changing the pH. These shifts are discussed in terms of a pH dependence of the haem midpoint redox potentials. The physiological relevance of this pH dependence is discussed.     

Burrowing beetles of the genus Bledius (Staphylinidae) as agents of bioturbation in the emergent areas and shores of an athalassic inland lake (Fuente de Piedra,southern of Spain)

Bledius (Elbidus) bicornis (Germ.) and B. (Eucerotobledius) furcatus (Oliv.) (Coleoptera: Staphylinidae) are the most important burrowing species in the emergent areas and shores in the athalassic lake of Fuente de Piedra (Málaga, S. of Spain). A first estimate of the importance of these organisms in this system is presented. These insects kick out sediment during their burrowing activity, which accumulates on the surface near the burrows as tumuli which can be easily eroded. The lake perimeter (17 km) is densely colonized (usual densities from 1700 to 2500 ind m⁻²). The amount of granulated material that can be potentially kicked out was 46.22 g dry wt m⁻² day⁻¹. At the same time, the material that constitutes the tumuli shows different characteristics from the compact ground below the surface. Thus, it is relatively enriched with organic matter (6.15 g per square meter), soluble phosphate (406.5 μg m⁻²) and ammonium (4856 μg m⁻²), whereas it lacks nitrate. Results of a transect from uninhabited areas to zones of maximum population density also show a similarity between the higher ground level of ammonium and phosphate concentrations and population density.     

Influence of temperature and salinity on carbon and nitrogen content in Dunaliella viridis teodoresco under nitrogen sufficiency

Cell carbon and nitrogen in D. viridis are strongly dependent on the culturing conditions. Both elements increase with increasing salinity. At 31°C cell carbon is maximum and cell nitrogen minimum. This temperature was described previously (Jiménez, C., Niell, F. X. & Fernandez, J. A. (1990). Hydrobiologia, 197, 165-72) as the optimal one for achieving the maximum oxygen evolution. These results point out a possible competence for the reducing power during carbon and nitrogen assimilation processes, and under conditions of high photosynthesis (carbon assimilation) there is a partial inhibition of nitrate reduction, making C:N ratio maximum under conditions of maximum net photosynthesis.The study of cell glycerol, nitrate, structural proteins and free amino acids indicates that all of these solutes accumulate in the cells as a result of the high salinity adaptation.     

Oxidation-reduction potentials of the hemes in cytochrome C3 from Desulfovibrio gigas in the presence and absence of ferredoxin by EPR spectroscopy.

1. Ferricytochrome c3 from D. gigas exhibits two low-spin ferric heme EPR resonances with gz-values at 2.959 and 2.853. Ferrocytochrome c3 is diamagnetic based on the absence of any EPR signals. 2. EPR potentiometric titrations result in the resolution of the two low-spin ferric heme resonances into two additional heme components representing in total the four hemes of the cytochrome, with EM values of -235 mV and -315 mV at heme resonance I and EM values of -235 mV and -306 mV at heme resonance II. 3. EPR spectroscopy has detected a significant diminution of intensity (approx. 60 p. 100) in the gx amplitude of ferricytochrome c3 in the presence of D. gigas ferredoxin II. The presence of ferredoxin II also causes a more negative shift in the EM of the second components of the signals at heme resonances I and II of cytochrome C3. Both observations suggest that an interaction has occurred between cytochrome C3 and ferredoxin II. 4. The results presented suggest that the heme ligand environment of ferricytochrome c3 from D. gigas is less perturbed and/or less asymmetric than environment for ferricytochrome c3 from D. vulgaris whose EPR behavior indicates the non-equivalence of all four hemes.     

Purification,characterization and biological activity of three forms of ferredoxin from the sulfate-reducing bacterium Desulfovibrio gigas

Three forms of ferredoxin FdI, FdI′, and FdII have been isolated from Desulfovibrio gigas, a sulfate reducer. They are separated by a combination of DEAE-cellulose and gel filtration chromatographic procedures. FdI and FdI′ present a slight difference in isoelectric point which enables the separation of the two forms over DEAE-cellulose, while FdII is easily separated from the two other forms by gel filtration. The three forms have the same amino acid composition and are isolated in different aggregation states. Molecular weight determinations by gel filtration gave values of 18 000 for FdI and FdI′ and 24 000 for FdII, whereas a value of 6000 is determined when dissociation is accomplished with sodium dodecyl sulfate. The electronic spectra are different and their ultraviolet-visible absorbance rations are 0.77, 0.87 and 0.68 respectively for FdI, FdI′ and FdII. Despite these differences, the physiological activities of the three forms are similar as far as the reduction of sulfite by molecular hydrogen is concerned.