128 research outputs found

    Dephosphorylation of the Thylakoid Membrane Light-Harvesting Complex-II by a Stromal Protein Phosphatase

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    Light-harvesting complex-II (LHC-II) phosphatase activity has generally been examined in the intact thylakoid membrane. A recent report of peptide-phosphatase activity associated with the chloroplast stromal fraction (Hammer, M.E et al. (1995) Photosynth Res 44:107–115) has led to the question of whether this activity is capable of dephosphorylating membrane-bound LHC-II. To this end, heat-treated thylakoid membranes were examined as a potential LHC-II phosphatase substrate. Following incubation of the thylakoid membrane at 60 °C for 15 min, the endogenous protein phosphatase and kinase activities were almost eliminated. Heat-inactivated phosphomembranes exhibited minimal dephosphorylation of the light harvesting complex-II. Peptide-phosphatase activities isolated from the thylakoid and stromal fraction were able to dephosphorylate LHC-II in heat-inactivated phosphomembranes. The stromal phosphatase showed highest activity against LHC-II at pH 9. Dephosphorylation of the LHC-II by the stromal enzyme was not inhibited by molybdate, vanadate or tungstate ions, but was partially inhibited by EDTA and a synthetic phosphopeptide mimicking the LHC-II phosphorylation site. Thus, the previously identified stromal phosphatase does appear capable of dephosphorylating authentic LHC-II in vivo

    Development of a System for Directed Evolution of \u3cem\u3eArabidopsis\u3c/em\u3e Formate Dehydrogenase to Utilize NADP as a Cofactor

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    Formate dehydrogenase (FDH) is a NAD-dependent enzyme found in methylotrophic bacteria, yeast and plants. This enzyme catalyzes the reversible oxidation of formate to carbon dioxide. The goal of this research was to determine the feasibility of using a directed evolution approach to generate an altered Arabidopsis FDH with a high affinity for NADP as a cofactor. A PCR procedure that induced approximately 1.5 mutations in the wild-type Arabidopsis FDH sequence per thousand base pairs was developed and the amplified products were transformed into E. coli cells. Approximately 1300 cell lines were assayed in 96-well microplates for activity with NADP+ and 100 putative mutants were selected for further study. One particular mutant line, pFDH-18, possessed reproducible NADP+-FDH activity. Sequence analysis showed that a single T in the wild-type DNA sequence had been changed to a G. The result of this mutation was that an isoleucine (Ile) residue at position 188 in the wild-type enzyme was converted to a methionine. This particular Ile residue is conserved in the known FDH sequences from higher plants and is located in the region of the enzyme that contains the binding domain for the NAD cofactor

    Corn Yield Potential and Optimal Soil Productivity in Irrigated Corn/Soybean Systems

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    In 1999, an interdisciplinary research team at the University of Nebraska established a field experiment to (1) quantify and understand the yield potential of corn and soybean under irrigated conditions, (2) identify efficient crop management practices to achieve yields that approach potential levels, and (3) determine the energy use efficiency, global warming and soil C-sequestration potential of intensively managed corn systems. The experiment compares systems that represent different levels of management intensity expressed as combinations of crop rotation (continuous corn, corn-soybean), plant density (low, medium, high) and nutrient management (recommended best management vs. intensive management). Detailed measurements include soil nutrient dynamics and C balance, crop growth and development, nutrient uptake and components of yield of corn and soybean, radiation use efficiency, soil surface fluxes of greenhouse gases, root biomass, C inputs through crop residues, translocation of non-structural carbohydrates, and amount, composition and activity of the microbial biomass. Selected results for corn are presented

    Corn Yield Potential and Optimal Soil Productivity in Irrigated Corn/Soybean Systems

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    In 1999, an interdisciplinary research team at the University of Nebraska established a field experiment to (1) quantify and understand the yield potential of corn and soybean under irrigated conditions, (2) identify efficient crop management practices to achieve yields that approach potential levels, and (3) determine the energy use efficiency, global warming and soil C-sequestration potential of intensively managed corn systems. The experiment compares systems that represent different levels of management intensity expressed as combinations of crop rotation (continuous corn, corn-soybean), plant density (low, medium, high) and nutrient management (recommended best management vs. intensive management). Detailed measurements include soil nutrient dynamics and C balance, crop growth and development, nutrient uptake and components of yield of corn and soybean, radiation use efficiency, soil surface fluxes of greenhouse gases, root biomass, C inputs through crop residues, translocation of non-structural carbohydrates, and amount, composition and activity of the microbial biomass. Selected results for corn are presented

    An Agent-Based Model to study the epidemiological and evolutionary dynamics of Influenza viruses

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    <p>Abstract</p> <p>Background</p> <p>Influenza A viruses exhibit complex epidemiological patterns in a number of mammalian and avian hosts. Understanding transmission of these viruses necessitates taking into account their evolution, which represents a challenge for developing mathematical models. This is because the phrasing of multi-strain systems in terms of traditional compartmental ODE models either requires simplifying assumptions to be made that overlook important evolutionary processes, or leads to complex dynamical systems that are too cumbersome to analyse.</p> <p>Results</p> <p>Here, we develop an Individual-Based Model (IBM) in order to address simultaneously the ecology, epidemiology and evolution of strain-polymorphic pathogens, using Influenza A viruses as an illustrative example.</p> <p>Conclusions</p> <p>We carry out careful validation of our IBM against comparable mathematical models to demonstrate the robustness of our algorithm and the sound basis for this novel framework. We discuss how this new approach can give critical insights in the study of influenza evolution.</p

    An Abundant Dysfunctional Apolipoprotein A1 in Human Atheroma

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    Recent studies have indicated that high-density lipoproteins (HDLs) and their major structural protein, apolipoprotein A1 (apoA1), recovered from human atheroma are dysfunctional and are extensively oxidized by myeloperoxidase (MPO). In vitro oxidation of either apoA1 or HDL particles by MPO impairs their cholesterol acceptor function. Here, using phage display affinity maturation, we developed a high-affinity monoclonal antibody that specifically recognizes both apoA1 and HDL that have been modified by the MPO-H2O2-Cl− system. An oxindolyl alanine (2-OH-Trp) moiety at Trp72 of apoA1 is the immunogenic epitope. Mutagenesis studies confirmed a critical role for apoA1 Trp72 in MPO-mediated inhibition of the ATP-binding cassette transporter A1 (ABCA1)-dependent cholesterol acceptor activity of apoA1 in vitro and in vivo. ApoA1 containing a 2-OH-Trp72 group (oxTrp72-apoA1) is in low abundance within the circulation but accounts for 20% of the apoA1 in atherosclerosis-laden arteries. OxTrp72-apoA1 recovered from human atheroma or plasma is lipid poor, virtually devoid of cholesterol acceptor activity and demonstrated both a potent proinflammatory activity on endothelial cells and an impaired HDL biogenesis activity in vivo. Elevated oxTrp72-apoA1 levels in subjects presenting to a cardiology clinic (n = 627) were associated with increased cardiovascular disease risk. Circulating oxTrp72-apoA1 levels may serve as a way to monitor a proatherogenic process in the artery wall

    Stable Isotope Biogeochemistry of Seabird Guano Fertilization: Results from Growth Chamber Studies with Maize (Zea Mays)

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    Stable isotope analysis is being utilized with increasing regularity to examine a wide range of issues (diet, habitat use, migration) in ecology, geology, archaeology, and related disciplines. A crucial component to these studies is a thorough understanding of the range and causes of baseline isotopic variation, which is relatively poorly understood for nitrogen (δ(15)N). Animal excrement is known to impact plant δ(15)N values, but the effects of seabird guano have not been systematically studied from an agricultural or horticultural standpoint.This paper presents isotopic (δ(13)C and δ(15)N) and vital data for maize (Zea mays) fertilized with Peruvian seabird guano under controlled conditions. The level of (15)N enrichment in fertilized plants is very large, with δ(15)N values ranging between 25.5 and 44.7‰ depending on the tissue and amount of fertilizer applied; comparatively, control plant δ(15)N values ranged between -0.3 and 5.7‰. Intraplant and temporal variability in δ(15)N values were large, particularly for the guano-fertilized plants, which can be attributed to changes in the availability of guano-derived N over time, and the reliance of stored vs. absorbed N. Plant δ(13)C values were not significantly impacted by guano fertilization. High concentrations of seabird guano inhibited maize germination and maize growth. Moreover, high levels of seabird guano greatly impacted the N metabolism of the plants, resulting in significantly higher tissue N content, particularly in the stalk.The results presented in this study demonstrate the very large impact of seabird guano on maize δ(15)N values. The use of seabird guano as a fertilizer can thus be traced using stable isotope analysis in food chemistry applications (certification of organic inputs). Furthermore, the fertilization of maize with seabird guano creates an isotopic signature very similar to a high-trophic level marine resource, which must be considered when interpreting isotopic data from archaeological material
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