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Human sulfite oxidase (hSO) was immobilised on SAM-coated silver electrodes under preservation of the native heme pocket structure of the cytochrome b5 (Cyt b5) domain and the functionality of the enzyme. The redox properties and catalytic activity of the entire enzyme were studied by surface enhanced resonance Raman (SERR) spectroscopy and cyclic voltammetry (CV) and compared to the isolated heme domain when possible. It is shown that heterogeneous electron transfer and catalytic activity of hSO sensitively depend on the local environment of the enzyme. Increasing the ionic strength of the buffer solution leads to an increase of the heterogeneous electron transfer rate from 17 s(-1) to 440 s(- 1) for hSO as determined by SERR spectroscopy. CV measurements demonstrate an increase of the apparent turnover rate for the immobilised hSO from 0.85 s(-1) in 100 mM buffer to 5.26 s(-1) in 750 mM buffer. We suggest that both effects originate from the increased mobility of the surface-bound enzyme with increasing ionic strength. In agreement with surface potential calculations we propose that at high ionic strength the enzyme is immobilised via the dimerisation domain to the SAM surface. The flexible loop region connecting the Moco and the Cyt b5 domain allows alternating contact with the Moco interaction site and the SAM surface, thereby promoting the sequential intramolecular and heterogeneous electron transfer from Moco via Cyt b5 to the electrode. At lower ionic strength, the contact time of the Cyt b5 domain with the SAM surface is longer, corresponding to a slower overall electron transfer process.
Plant outward-rectifying K+ channels mediate K+ efflux from guard cells during stomatal closure and from root cells into the xylem for root-shoot allocation of potassium (K). Intriguingly, the gating of these channels depends on the extracellular K+ concentration, although the ions carrying the current are derived from inside the cell. This K+ dependence confers a sensitivity to the extracellular K+ concentration ([K+]) that ensures that the channels mediate K+ efflux only, regardless of the [K+] prevailing outside. We investigated the mechanism of K+-dependent gating of the K+ channel SKOR of Arabidopsis by site-directed mutagenesis. Mutations affecting the intrinsic K+ dependence of gating were found to cluster in the pore and within the sixth transmembrane helix (S6), identifying an 'S6 gating domain' deep within the membrane. Mapping the SKOR sequence to the crystal structure of the voltage-dependent K+ channel KvAP from Aeropyrum pernix suggested interaction between the S6 gating domain and the base of the pore helix, a prediction supported by mutations at this site. These results offer a unique insight into the molecular basis for a physiologically important K+-sensory process in plants
The Mexican tetra Astyanax fasciatus has evolved a variety of more or less color- and eyeless cave populations. Here we examined the evolution of the female preference for large male body size within different populations of this species, either surface- or cave-dwelling. Given the choice between visual cues from a large and a small male, females from the surface form as well as females from an eyed cave form showed a strong preference for large males. When only non-visual cues were presented in darkness, the surface females did not prefer either males. Among the six cave populations studied, females of the eyed cave form and females of one of the five eyeless cave populations showed a preference for large males. Apparently, not all cave populations of Astyanax have evolved non-visual mating preferences. We discuss the role of selection by benefits of non-visual mate choice for the evolution of non-visual mating preferences
Filamin c is the predominantly expressed filamin isoform in striated muscles. It is localized in myofibrillar Z- discs, where it binds FATZ and myotilin, and in myotendinous junctions and intercalated discs. Here, we identify Xin, the protein encoded by the human gene 'cardiomyopathy associated 1' (CMYA1) as filamin c binding partner at these specialized structures where the ends of myofibrils are attached to the sarcolemma. Xin directly binds the EVH1 domain proteins Mena and VASP. In the adult heart, Xin and Mena/VASP colocalize with filamin c in intercalated discs. In cultured cardiomyocytes, the proteins also localize in the nonstriated part of myofibrils, where sarcomeres are assembled and an extensive reorganization of the actin cytoskeleton occurs. Unusual intraexonic splicing events result in the existence of three Xin isoforms that associate differentially with its ligands. The identification of the complex filamin c-Xin-Mena/VASP provides a first glance on the role of Xin in the molecular mechanisms involved in developmental and adaptive remodeling of the actin cytoskeleton during cardiac morphogenesis and sarcomere assembly. (c) 2006 Elsevier Inc. All rights reserved
1.Interactions among plants are key processes that strongly influence the structure and dynamics of plant populations and communities. However, most empirical studies of plant-plant interactions failed to repeatedly measure the plants? response to neighboring individuals and thereby neglected possible changes in interactions throughout the life history of the plants.2.Here, we tested the hypothesis that competition between annual species intensifies from early to late life history stages. To test this hypothesis, we sequentially measured interactions at different levels of water stress. 3.For this purpose, we conducted neighbor-removal experiments in three study sites located along a climatic gradient in Israel. The two annual species Biscutella didyma and Hymenocarpos circinnatus were used as target plants. They grew with and without neighbors in their natural habitats. Five response variables, according to the consecutive life-history stages, (seedling survival, juvenile biomass, adult survival, number of seeds and final biomass) were recorded throughout the whole growing season. 4.The results suggest that direction and intensity of interactions varied considerably between environments and life stages. On average, growth-related response variables indicated higher competition intensity at the productive end of the climatic gradient, while survival indicated either facilitation at the dry end or no trend along the gradient. 5.Considering the temporal aspect, moderate facilitation short after germination shifted to strong competition at the end of the growing season. 6.Our results highlight that the outcome of experimental studies on plant-plant interactions may not only depend on the environmental productivity but even more on the life stage at which a target plant is found.
In semiarid savannas of Southern Africa current land use practices and climate change may lead to substantial changes of vegetation structure in the near future, however uncertainty remains about the potential consequences and the magnitude of change. In this paper we study the impact of climate change, cattle grazing, and wood cutting on shrub cover dynamics in savannas of the southern Kalahari. We use an established savanna ecosystem model to simulate landscape dynamics in terms of rainfall, fire and distribution of the dominant tree Acacia erioloba. We then incorporate these data into a spatial population model of the common, fleshy-fruited shrub Grewia flava and investigate shrub cover dynamics for a period of 100 years. Depending on the intensity of commercial wood cutting practices tree removal of A. erioloba led to a strong decline of the G. flava population, as shrub recruitment is concentrated in tree sub-canopies due to bird-mediated seed dispersal. Under climate change shrub cover slightly decreased with decreasing precipitation and was unchanged with increase in precipitation variability. Contrarily, grazing by cattle strongly increased shrub cover and facilitated shrub encroachment because of cattle-induced distribution of G. flava seeds into the matrix vegetation. Knowledge of the latter process is particularly important because shrub invasion is a major concern for conservation and savanna rangeland management as a result of its adverse effects on livestock carrying capacity and biodiversity
Use of large Acacia trees by the cavity dwelling Black-tailed Tree Rat in the southern Kalahari
(2006)
Recent extensive harvesting of large, often dead Acacia trees in and savanna of southern Africa is cause for concern about the conservation status of the arid savanna and its animal community. We mapped vegetation and nests of the Black-tailed Tree Rat Thallomy's nigricauda to assess the extent to which the rats depend on particular tree species and on the existence of dead, standing trees. The study was conducted in continuous Acacia woodland on the southern and eastern edge of the Kalahari, South Africa. Trees in which there were tree rat nests were compared with trees of similar size and vigour to identify the characteristics of nest sites. Spatial analysis of tree rat distribution was conducted using Ripley's-L function. We found that T nigricauda was able to utilize all available tree species, as long as trees were large and old enough so that cavities were existing inside the stem. The spatial distribution of nest trees did not show clumping at the investigated scale, and we therefore reject the notion of the rats forming colonies when inhabiting continuous woodlands. The selection of a particular tree as a nest site was furthermore depending on the close proximity of the major food plant, Acacia mellifera. This may limit the choice of suitable nest sites. since A. mellifera was less likely to grow within a vegetation patch containing a large trees than in patches without large trees.
Representatives of the genus Stentor (Stentoridae, Heterotrichea) are striking ciliates in environmental water samples because of their size (up to 4 mm) and their trumpet-like shape. Important for species identification are the following main characteristics: (1) the presence or absence of endosymbiotic algae (zoochlorellae); (2) the colour of the pigmented cortical granules, and (3) the shape of the macronucleus. The complete small subunit rDNA (SSU rDNA) of 19 further representatives of the genus Stentor was sequenced to examine the phylogenetic relationships within this genus and to determine the taxonomic value of these main characteristics. The detailed phylogenetic analyses yielded a separation of all species possessing a single compact macronucleus from those species with an "elongated" macronucleus (moniliform or vermiform). The data also indicate that the uptake of algae as well as the loss of pigmentation happened independently in different lineages. Furthermore, a high level of intraspecific variation within several species was found. Thus, S. muelleri and S. (sp.) cf. katashimai appear to represent distinct species and S. multiformis is composed of a species complex.
Tight junctions seal intercellular clefts via membrane-related strands, hence, maintaining important organ functions. We investigated the self-association of strand-forming transmembrane tight junction proteins. The regulatory tight junction protein occludin was differently tagged and cotransfected in eucaryotic cells. These occludins colocalized within the plasma membrane of the same cell, coprecipitated and exhibited fluorescence resonance energy transfer. Differently tagged strand-forming claudin-5 also colocalized in the plasma membrane of the same cell and showed fluorescence resonance energy transfer. This demonstrates self-association in intact cells both of occludin and claudin-5 in one plasma membrane. In search of dimerizing regions of occludin, dimerization of its cytosolic C-terminal coiled-coil domain was identified. In claudin-5, the second extracellular loop was detected as a dimer. Since the transmembrane junctional adhesion molecule also is known to dimerize, the assumption that homodimerization of transmembrane tight junction proteins may serve as a common structural feature in tight junction assembly is supported
The nature of the periplastidial pathway of starch biosynthesis was investigated with the model cryptophyte Guillardia theta. The storage polysaccharide granules were shown to be composed of both amylose and amylopectin fractions with a chain length distribution and crystalline organization very similar to those of starch from green algae and land plants. Most starch granules displayed a shape consistent with biosynthesis occurring around the pyrenoid through the rhodoplast membranes. A protein with significant similarity to the amylose-synthesizing granule-bound starch syntbase 1 from green plants was found as the major polypeptide bound to the polysaccharide matrix. N-terminal sequencing of the mature protein proved that the precursor protein carries a nonfunctional transit peptide in its bipartite topogenic signal sequence which is cleaved without yielding transport of the enzyme across the two inner plastid membranes. The enzyme was shown to display similar affinities for ADP and UDP-glucose, while the V-max measured with UDP-glucose was twofold higher. The granule-bound starch synthase from Guillardia theta was demonstrated to be responsible for the synthesis of long glucan chains and therefore to be the functional equivalent of the amylose- synthesizing enzyme of green plants. Preliminary characterization of the starch pathway suggests that Guillardia theta utilizes a UDP-glucose-based pathway to synthesize starch