The "prismane" protein resolved: X-ray structure at 1.7 Å and multiple spectroscopy of two novel 4Fe clusters

The three-dimensional structure of the native "putative prismane" protein from Desulfovibrio vulgaris (Hildenborough) has been solved by X-ray crystallography to a resolution of 1.72?Å. The molecule does not contain a [6Fe-6S] prismane cluster, but rather two 4Fe clusters some 12?Å apart and situated close to the interfaces formed by the three domains of the protein. Cluster 1 is a conventional [4Fe-4S] cubane bound, however, near the N-terminus by an unusual, sequential arrangement of four cysteine residues (Cys 3, 6, 15, 21). Cluster 2 is a novel 4Fe structure with two μ₂-sulfido bridges, two μ₂-oxo bridges, and a partially occupied, unidentified μ₂ bridge X. The protein ligands of cluster 2 are widely scattered through the second half of the sequence and include three cysteine residues (Cys 312, 434, 459), one persulfido-cysteine (Cys 406), two glutamates (Glu 268, 494), and one histidine (His 244). With this unusual mixture of bridging and external type of ligands, cluster 2 is named the "hybrid" cluster, and its asymmetric, open structure suggests that it could be the site of a catalytic activity. X-ray absorption spectroscopy at the Fe K-edge is readily interpretable in terms of the crystallographic model when allowance is made for volume contraction at 10?K; no Fe··Fe distances beyond 3.1?Å could be identified. EPR, Mössbauer and MCD spectroscopy have been used to define the oxidation states and the magnetism of the clusters in relation to the crystallographic structure. Reduced cluster 1 is a [4Fe-4S]¹⁺ cubane with S?=?3/2; it is the first biological example of a "spin-admixed" iron-sulfur cluster. The hybrid cluster 2 has four oxidation states from (formally) all Fe^III to three Fe^II plus one Fe^III. The four iron ions are exchange coupled resulting in the system spins S?=?0, 9/2, 0 (and 4), 1/2, respectively, for the four redox states. Resonance Raman spectroscopy suggests that the bridging ligand X which could not be identified unambiguously in the crystal structure is a solvent-exchangeable oxygen.     

Seedling establishment in different microsites on Mount St. Helens,Washington, USA

On the volcanically devastated Pumice Plain of Mount St. Helens, plant species colonized microsites differentially. Peak colonization did not occur in the same microsites as peak establishment and growth. In addition, observed microsite colonization patterns differed between years. Two studies were conducted. The first assessed seedling establishment and growth from seeds sown at different microsites. The second assessed colonization into four microsite types that were constructed on the Pumice Plain. Hypochaeris radicata was the most common species to survive when the same number of seeds of four species were planted; however, Anaphalis margaritacea was the most common colonizer of microsites. Microsites with the largest biomass plants in the first study generally had the highest colonization in the second study. Sites that do not possess features to trap seeds, such as flats and ridges, are not opportune places for a plant to grow since there is little microclimatic or substrate amelioration. Thus, flat microsites had low biomass in the establishment experiment due to the lack of amelioration and contained few plants in the colonization experiment due to a lack of seed trapping mechanisms. These results show that in the primary successional landscape of Mount St. Helens microsites are critical to revegetation dynamics. Changes in the pattern of microsite colonization between years emphasizes the dynamic nature of the landscape and the important influences of climate, substrate amelioration and seed rain to plant establishment.     

Simultaneous analysis of multiple aminothiols in human plasma by high performance liquid chromatography with fluorescence detection

Aminothiols serve numerous vital functions in biochemistry, including detoxification and regulation of cellular metabolism, enzymatic activity, and protein trafficking and degradation. Plasma aminothiol concentrations are frequently measured for clinical and translational research investigating oxidative stress, and for routine clinical diagnosis and monitoring of vascular injury. Although a variety of techniques are available to measure aminothiol concentrations in plasma, high performance liquid chromatography with fluorescence detection (HPLC–FD) is the most widely used. This review summarizes HPLC–FD methods, including pre-analytical considerations, procedures for sample reduction, derivatization, and chromatographic separation of the primary biological aminothiols cysteine, homocysteine, cysteinylglycine, and glutathione in human plasma.     

REPRODUCTIVE OUTPUT AND BIOMASS ALLOCATION IN SESBANIA EMERUS IN A TROPICAL SWAMP

This study examined the correlation of moisture, reproductive phenology, density of mature plants, and herbivory of apical meristems with the morphology and reproductive output of Sesbania emerus, an annual legume growing along a moisture gradient in a swamp in Guanacaste Province, Costa Rica. It also determined how biomass allocation varied and how it was related to reproductive output of plants growing along the moisture gradient within the swamp. Morphological changes included production of more stems and branches in response to herbivory, more and higher prop roots and more aerenchyma as water depth increased, and greater stem diameter in lower densities. Plant height varied greatly within a site, but not among habitats. Plants began to produce flowers and fruits later in wet sites than in dry sites. Reproductive output was generally more sensitive to environmental variables than was plant size. Fruit number and plant height were positively correlated for almost every treatment. Greater fruit and seed production were correlated with drier sites, earlier phenology, and lower density, but not with herbivory. Total biomass accumulation did not vary among moisture sites, but root production appeared to occur at the expense of reproductive output in the wetter sites. Plants in the wetter sites had both a greater percent and a greater absolute amount of biomass in roots, and a lower percent and a lower absolute amount of biomass in fruits and seeds than plants in drier sites. The root: shoot ratio was nearly five times higher in the wet than the dry site. Seed number per plant ranged from a mean of 6,800 at the wet site to a mean of 16,878 at the dry site. If this striking phenotypic variation in reproductive output and biomass accumulation has a genetic basis, the possibility of ecotypic differentiation exists in S. emerus.     

Arabinan Metabolism during Seed Development and Germination in Arabidopsis

Arabinans are found in the pectic network of many cell walls, where, along with galactan, they are present as side chains of Rhamnogalacturonan I. Whilst arabinans have been reported to be abundant polymers in the cell walls of seeds from a range of plant species, their proposed role as a storage reserve has not been thoroughly investigated. In the cell walls of Arabidopsis seeds, arabinose accounts for approximately 40% of the monosaccharide composition of non- cellulosic polysaccharides of embryos. Arabinose levels decline to -15% during seedling establishment, indicating that cell wall arabinans may be mobilized during germination. Immunolocalization of arabinan in embryos, seeds, and seedlings reveals that arabinans accumulate in developing and mature embryos, but disappear during germination and seedling establishment. Experiments using 14C-arabinose show that it is readily incorporated and metabolized in growing seedlings, indicating an active catabolic pathway for this sugar. We found that depleting arabinans in seeds using a fungal arabinanase causes delayed seedling growth, lending support to the hypothesis that these polymers may help fuel early seedling growth.