Ferritin-catalyzed consumption of hydrogen peroxide by amine buffers causes the variable Fe2+ to O2 stoichiometry of iron deposition in horse spleen ferritin

Ferritin catalyzes the oxidation of Fe2+ by O2 to form a reconstituted Fe3+ oxy-hydroxide mineral core, but extensive studies have shown that the Fe2+ to O2 stoichiometry changes with experimental conditions. At Fe2+ to horse spleen ferritin (HoSF) ratios greater than 200, an upper limit of Fe2+ to O2 of 4 is typically measured, indicating O2 is reduced to 2H2O. In contrast, a lower limit of Fe2+ to O2 of approximately 2 is measured at low Fe2+ to HoSF ratios, implicating H2O2 as a product of Fe2+ deposition. Stoichiometric amounts of H2O2 have not been measured, and H2O2 is proposed to react with an unknown system component. Evidence is presented that identifies this component as amine buffers, including 3-N-morpholinopropanesulfonic acid (MOPS), which is widely used in ferritin studies. In the presence of non-amine buffers, the Fe2+ to O2 stoichiometry was approximately 4.0, but at high concentrations of amine buffers (0.10 M) the Fe2+ to O2 stoichiometry is approximately 2.5 for iron loadings of eight to 30 Fe2+ per HoSF. Decreasing the concentration of amine buffer to zero resulted in an Fe2+ to O2 stoichiometry of approximately 4. Direct evidence for amine buffer modification during Fe2+ deposition was obtained by comparing authentic and modified buffers using mass spectrometry, NMR, and thin layer chromatography. Tris(hydroxymethyl)aminomethane, MOPS, and N-methylmorpholine (a MOPS analog) were all rapidly chemically modified during Fe2+ deposition to form N-oxides. Under identical conditions no modification was detected when amine buffer, H2O2, and O2 were combined with Fe2+ or ferritin separately. Thus, a short-lived ferritin intermediate is required for buffer modification by H2O2. Variation of the Fe2+ to O2 stoichiometry versus the Fe2+ to HoSF ratio and the amine buffer concentration are consistent with buffer modification.     

鄱阳湖湿地两种优势植物叶片C、N、P动态特征

2011年2—6月在鄱阳湖南矶湿地国家级自然保护区逐月测定了灰化苔草(Carex cinerascens)、南荻(Triarrhena lutarioriparia)叶片C、N、P含量及其地上生物量,以阐明鄱阳湖湿地优势植物C、N、P含量及化学计量比动态特征与控制因子,探讨湿地养分利用与限制状况。结果表明：1）两种优势植物叶有机碳含量变化范围分别为365.3—386.6 mg/g和352.6—393.2 mg/g,平均值(?标准差)分别为(375.5?17.4) mg/g和(371.7?12.5) mg/g;叶N含量分别为6.96—17.59 mg/g和5.50—20.68 mg/g,平均值分别为(11.35?1.40) mg/g和(11.54?0.84) mg/g;叶P含量变化范围为0.65—2.14 mg/g和0.57—2.25 mg/g,平均含量为(1.56?0.69) mg/g和(1.55?0.68) mg/g。两种植物C:N、C:P、N:P平均值分别为37.65、413.60、9.62和41.05、410.29、9.57,C、N、P及其化学计量比种间差异不显著(P>0.05)。2）气温与地上生物量是N、P及其化学计量比季节变化的主要控制因子,气温和生物量对两种优势植物叶片氮、磷含量的影响要高于对叶有机碳含量的影响。3）植物C:N、C:P与地上生物量变化趋势基本一致,显示N、P养分利用效率随植物的快速生长而提高;根据两种优势植物及土壤N、P含量与化学计量比来判断,研究区植物更多地受氮限制。     

Interactions of Potent Multisubstrate Analogue Inhibitors with Purine Nucleoside Phosphorylase from Calf Spleen—Kinetic and Spectrofluorimetric Studies

Abstract

Dissociation constants and stoichiometry of binding for interaction of trimeric calf spleen purine nucleoside phosphorylase with potent multisubstrate analogue inhibitors were studied by kinetic and spectrofluorimetric methods.     

Modeling the bacterial contribution to planktonic community respiration in the regulation of solar energy and nutrient availability

In planktonic ecosystems, algae and bacteria exhibit complex interrelationships, as algae provide an important organic matter source for microbial growth while microbial metabolism recycles limiting nutrients for algae in a loose commensalism. However, algae and bacteria can also compete for available nutrients if supplies of organic matter are sufficient to satisfy bacterial demand. We developed a stoichiometrically explicit model of bacteria–algae interactions that incorporated realistic assumptions about algal light and nutrient utilization, algal exudation of organic matter, and bacterial processing of organic matter and nutrients. The model makes specific predictions about how the relative balance of algae and bacteria should change in response to varied nutrient and light availability seen in lakes and oceans. The model successfully reproduces published empirical data and indicates that, under moderate nutrient supply, the bacterial percentage of total respiration should be maximal at intermediate light intensity.     

New dimensions in the study of protein complexes using quantitative mass spectrometry

Mass spectrometry combined with affinity purification techniques has evolved as a prime tool for the in-depth study of distinct protein complexes and protein-protein interactions. It fueled proteome-wide studies leading to the establishment of intricate cellular protein interaction networks. Recent innovative advancements in quantitative protein mass spectrometry act as driving force for the design of ingenious strategies in interaction proteomics facilitating the acquisition of interaction data with improved accuracy and, most intriguingly, the elucidation of functional aspects by monitoring transient interactions as well as dynamic changes in composition, stoichiometry, localization and post-translational modification of protein complexes under various conditions.     

不同潮汐和盐度下红树植物幼苗秋茄的化学计量特征

借助室内潮汐模拟系统,分析秋茄幼苗的叶片、茎,根在不同淹浸时间梯度和盐度梯度下N、P、K、Ca、Cl、Mg、Al、Fe、Zn、Cu、B等元素的化学计量特征变化.结果表明水文因子能够显著影响秋茄不同器官的化学计量特征;幼苗的元素化学计量在盐度和淹浸两类水文因子下表现出不同的响应特征:特别是N、K、Ca、Cl、Mg、Na、Zn,Cu元素含量在不同程度的水文胁迫下存在明显的差异;盐度组内,幼苗对Na,Cl的选择性吸收随盐度增高而增强,同时对K、Ca,Mg的吸收减弱;随盐度的增大,盐分(Na、Cl、K、Ca、Mg)在不同器官中的含量表现出趋同效应;淹浸组N:P比为3-11,盐度组N:P比为6-15,明显小于同类研究.综合分析认为秋茄幼苗受到N元素限制,而P元素始终未成为限制因子,随水文胁迫的加强,N限制也加大,且盐度胁迫比淹浸胁迫表现的更明显.通过化学计量学和水文学的分析,为摸清中国红树林保护和恢复的主要红树植物秋茄的适生环境提供科学依据.     

A Kunitz proteinase inhibitor from Archidendron ellipticum seeds: purification, characterization, and kinetic properties

Leguminous plants in the tropical rainforests are a rich source of proteinase inhibitors and this work illustrates isolation of a serine proteinase inhibitor from the seeds of Archidendron ellipticum (AeTI), inhabiting Great Nicobar Island, India. AeTI was purified to homogeneity by acetone and ammonium sulfate fractionation, and ion exchange, size exclusion and reverse phase chromatography (HPLC). SDS-PAGE of AeTI revealed that it is constituted by two polypeptide chains (alpha-chain, M(r) 15,000 and beta-chain, M(r) 5000), the molecular weight being approximately 20 kDa. N-terminal sequence showed high homology with other serine proteinase inhibitors belonging to the Mimosoideae subfamily. Both Native-PAGE as well as isoelectric focussing showed four isoinhibitors (pI values of 4.1, 4.55, 5.27 and 5.65). Inhibitory activity of AeTI remained unchanged over a wide range of temperatures (0-60 degrees C) and pH (1-10). The protein inhibited trypsin in the stoichiometric ratio of 1:1, but lacked similar stoichiometry against chymotrypsin. Also, AeTI-trypsin complex was stable to SDS unlike the SDS unstable AeTI-chymotrypsin complex. AeTI, which possessed inhibition constants (K(i)) of 2.46 x 10(-10) and 0.5 x 10(-10)M against trypsin and chymotrypsin activity, respectively, retained over 70% of inhibitory activity after being stored at -20 degrees C for more than a year. Initial studies on the insecticidal properties of AeTI indicate it to be a very potent insect anti-feedant.     

Simulation modeling of ligand receptor interactions at non-equilibrium conditions: processing of noisy inputs by ionotropic receptors

The first event in signal transduction at a synapse is the binding of transmitters to receptors. Because of rapidly changing transmitter levels this binding is unlikely to occur at equilibrium. We describe a mathematical approach that models complex receptor interactions in which the timing and amplitude of transmitter release are noisy. We show that exact solutions for simple bimolecular interactions and receptor transitions can be used to model complex reaction schemes by expressing them in sets of difference equations. Results from the difference equation method to describe binding and channel opening at extended time points compare well with standard solutions using ordinary differential equations. Because it is applicable to noisy systems we used the difference method to investigate the information processing capabilities of GABA receptors and predict how pharmacological agents may modify these properties. As previously demonstrated, the response to a single pulse of GABA is prolonged through entry into a desensitized state. During trains of stimuli the signal to noise ratio can change, and even increase progressively, but the overall transmitted fidelity of the signal decreases with increased driving frequency. The GABA modulator chlorpromazine (primarily affects agonist on and off rates) is predicated to increase receptor signal to noise ratio at all frequencies whereas pregnenolone sulfate (affects receptor desensitization) completely inhibits information transfer.     

Modeling and analysis of stoichiometric two-patch consumer-resource systems

Ecological stoichiometry studies the balance of energy and multiple chemical elements in ecological interactions to establish how the laws of thermodynamics affect food-web dynamics and nutrient cycling in ecosystems. In this paper, we incorporate stoichiometric principles in a model with habitat heterogeneity and dispersal in order to better understand population growth dynamics. This model describes a situation where a resource is separated into two patches by a barrier. Growth of the resource in each patch is limited by soil fertility and self-crowding. The consumer disperses between the two patches and is not affected by the barrier. The consumer's growth is potentially limited by the phosphorus content of the acquired resource. Mathematical analysis of this model and simulations are performed. Several biological implications, including an observed "stoichiometric extinction effect," are demonstrated with simulation where the stoichiometric mechanism appears to promote extinction in a patchy environment. This is in sharp contrast to the notion that stoichiometry mechanism promotes diversity in spatially homogeneous settings. Another important result is the rediscovery of a simple and plausible biological mechanism that generates local and global extinction. In this setting, which can be considered a spatially mediated form of apparent competition, the dispersal of the consumer from the rich patch can des the growth of the resource in the poor patch and in some situations can lead to the extinction of the resource in the poor patch.     

Stoichiometric differences in food quality: impacts on genetic diversity and the coexistence of aquatic herbivores in a <Emphasis Type="Italic">Daphnia</Emphasis> hybrid complex

The maintenance of genetic and species diversity in an assemblage of genotypes (clones) in the Daphnia pulex species complex (Cladocera: Anomopoda) in response to variation in the carbon:phosphorus ratio (quantity and quality) of the green alga, Scenedesmus acutus, was examined in a 90-day microcosm competition experiment. Results indicated that mixed assemblages of seven distinct genotypes (representing clonal lineages of D. pulex, D. pulicaria and interspecific hybrids) showed rapid loss of genetic diversity in all treatments (2 × 2 factorial design, high vs. low quantity, and high vs. low quality). However, the erosion of diversity (measured as the effective number of clones) was slowest under the poorest food conditions (i.e., low quantity, low quality) and by the conclusion of the experiment (90 days) had resulted in the (low, low) treatment having significantly greater genetic diversity than the other three treatments. In addition, significant genotype (clone) × (food) environment interactions were observed, with a different predominant species/clone found under low food quality versus high food quality (no significant differences were detected for the two food quantities). A clone of D. pulex displaced the other clones under low food quality conditions, while a clone of D. pulicaria displaced the other clones in the high food quality treatments. Subsequent life-history experiments were not sufficient to predict the outcome of competitive interactions among members of this clonal assemblage. Our results suggest that genetic diversity among herbivore species such as Daphnia may be impacted not only by differences in food quantity but also by those in food quality and could be important in the overall maintenance of genetic diversity in natural populations. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.