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991.
Cloning,Expression and Purification of an Acetoacetyl CoA Thiolase from Sunflower Cotyledon
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James H. Dyer Anthony Maina Iris D. Gomez Melissa Cadet Silke Oeljeklaus Anke C. Schiedel 《International journal of biological sciences》2009,5(7):736-744
Thiolase I and II coexist as part of the glyoxysomal β-oxidation system in sunflower (Helianthus annuus L.) cotyledons, the only system shown to have both forms. The importance of thiolases can be underscored not only by their ubiquity, but also by their involvement in a wide variety of processes in plants, animals and bacteria. Here we describe the cloning, expression and purification of acetoacetyl CoA thiolase (AACT) in enzymatically active form. Use of the extensive amount of sequence information from the databases facilitated the efficient generation of the gene-specific primers used in the RACE protocols. The recombinant AACT (1233 bp) shares 75% similarity with other plant AACTs. Comparison of specific activity of this recombinant AACT to a previously reported enzyme purified from primary sunflower cotyledon tissue was very similar (263 nkat/mg protein vs 220 nkat/mg protein, respectively). Combining the most pure fractions from the affinity column, the enzyme was purified 88-fold with a 55% yield of the enzymatically active, 47 kDa AACT. 相似文献
992.
The role of secondary heart field in cardiac development 总被引:1,自引:0,他引:1
Laura A. Dyer 《Developmental biology》2009,336(2):137-376
Although de la Cruz and colleagues showed as early as 1977 that the outflow tract was added after the heart tube formed, the source of these secondarily added cells was not identified for nearly 25 years. In 2001, three pivotal publications described a secondary or anterior heart field that contributed to the developing outflow tract. This review details the history of the heart field, the discovery and continuing elucidation of the secondarily adding myocardial cells, and how the different populations identified in 2001 are related to the more recent lineage tracing studies that defined the first and second myocardial heart fields/lineages. Much recent work has focused on secondary heart field progenitors that give rise to the myocardium and smooth muscle at the definitive arterial pole. These progenitors are the last to be added to the arterial pole and are particularly susceptible to abnormal development, leading to conotruncal malformations in children. The major signaling pathways (Wnt, BMP, FGF8, Notch, and Shh) that control various aspects of secondary heart field progenitor behavior are discussed. 相似文献
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995.
Lee DL Sturgis LC Labazi H Osborne JB Fleming C Pollock JS Manhiani M Imig JD Brands MW 《American journal of physiology. Heart and circulatory physiology》2006,290(3):H935-H940
Plasma levels of IL-6 correlate with high blood pressure under many circumstances, and ANG II has been shown to stimulate IL-6 production from various cell types. This study tested the role of IL-6 in mediating the hypertension caused by high-dose ANG II and a high-salt diet. Male C57BL6 and IL-6 knockout (IL-6 KO) mice were implanted with biotelemetry devices and placed in metabolic cages to measure mean arterial pressure (MAP), heart rate (HR), sodium balance, and urinary albumin excretion. Baseline MAP during the control period averaged 114 +/- 1 and 109 +/- 1 mmHg for wild-type (WT) and IL-6 KO mice, respectively, and did not change significantly when the mice were placed on a high-salt diet (HS; 4% NaCl). ANG II (90 ng/min sc) caused a rapid increase in MAP in both groups, to 141 +/- 9 and 141 +/- 4 in WT and KO mice, respectively, on day 2. MAP plateaued at this level in KO mice (134 +/- 2 mmHg on day 14 of ANG II) but began to increase further in WT mice by day 4, reaching an average of 160 +/- 4 mmHg from days 10 to 14 of ANG II. Urinary albumin excretion on day 4 of ANG II was not different between groups (9.18 +/- 4.34 and 8.53 +/- 2.85 microg/2 days for WT and KO mice). By day 14, albumin excretion was nearly fourfold greater in WT mice, but MAP dropped rapidly back to control levels in both groups when the ANG II was stopped after 14 days. Thus the approximately 30 mmHg greater ANG II hypertension in the WT mice suggests that IL-6 contributes significantly to ANG II-salt hypertension. In addition, the early separation in MAP, the albumin excretion data, and the rapid, post-ANG II recovery of MAP suggest an IL-6-dependent mechanism that is independent of renal injury. 相似文献
996.
Tung tree DGAT1 and DGAT2 have nonredundant functions in triacylglycerol biosynthesis and are localized to different subdomains of the endoplasmic reticulum 总被引:18,自引:0,他引:18
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Shockey JM Gidda SK Chapital DC Kuan JC Dhanoa PK Bland JM Rothstein SJ Mullen RT Dyer JM 《The Plant cell》2006,18(9):2294-2313
Seeds of the tung tree (Vernicia fordii) produce large quantities of triacylglycerols (TAGs) containing approximately 80% eleostearic acid, an unusual conjugated fatty acid. We present a comparative analysis of the genetic, functional, and cellular properties of tung type 1 and type 2 diacylglycerol acyltransferases (DGAT1 and DGAT2), two unrelated enzymes that catalyze the committed step in TAG biosynthesis. We show that both enzymes are encoded by single genes and that DGAT1 is expressed at similar levels in various organs, whereas DGAT2 is strongly induced in developing seeds at the onset of oil biosynthesis. Expression of DGAT1 and DGAT2 in yeast produced different types and proportions of TAGs containing eleostearic acid, with DGAT2 possessing an enhanced propensity for the synthesis of trieleostearin, the main component of tung oil. Both DGAT1 and DGAT2 are located in distinct, dynamic regions of the endoplasmic reticulum (ER), and surprisingly, these regions do not overlap. Furthermore, although both DGAT1 and DGAT2 contain a similar C-terminal pentapeptide ER retrieval motif, this motif alone is not sufficient for their localization to specific regions of the ER. These data suggest that DGAT1 and DGAT2 have nonredundant functions in plants and that the production of storage oils, including those containing unusual fatty acids, occurs in distinct ER subdomains. 相似文献
997.
Peter Michael Moyle Colleen Olive Levente Karpati Nadia Barozzi Mei-Fong Ho Joanne Dyer Hsien Kuo Sun Michael Good Istvan Toth 《International journal of peptide research and therapeutics》2006,12(3):317-326
Group A streptococcus (GAS) is responsible for causing many clinical complications including the relatively benign streptococcal pharyngitis and impetigo. However, if left untreated, these conditions may lead to more severe diseases such as rheumatic fever (RF) and rheumatic heart disease (RHD). These diseases exhibit high morbidity and mortality, particularly in developing countries and in indigenous populations of affluent countries. As RF and RHD only ever occur following GAS infection, a vaccine offers promise for their prevention. As such, we have investigated the use of the lipid-core peptide (LCP) system for the development of multi-valent prophylactic GAS vaccines. The current study has investigated the capacity of this system to adjuvant up to four different GAS peptide epitopes. Presented are the synthesis and immunological assessment of tetra-valent and tri-valent GAS LCP systems. We demonstrated their capacity to elicit systemic IgG antibody responses in B10.BR mice to all GAS peptide epitopes. The data also showed that the LCP systems were self-adjuvanting. These findings are particularly encouraging for the development of multi-valent LCP-based GAS vaccines. 相似文献
998.
Researchers examining the mechanisms of ichthyotoxicity of Pfiesteria shumwayae have come to different conclusions about the role of toxin in this process. Some attribute fish mortality solely to direct attack by these pedunculate dinoflagellates on exposed fish tissue while others have provided evidence for a role of a soluble toxin. Detection of toxin, especially in low concentrations, is a function of the sensitivity of the selected bioassay methods and the various groups addressing this question have utilized different methods. One notable difference in fish bioassay methods utilized to detect Pfiesteria-associated toxin (PfTx) is the species of fish tested. Studies that have not detected PfTx in bioassays generally have used Cyprinodon variegatus (sheepshead minnow) as the test fish while those that have detected toxin generally used Oreochromis spp. (Tilapia). In this study response of these two fish species was compared to determine their relative sensitivity to physical attack by P. shumwayae and to PfTx. The results indicate that Oreochromis niloticus is more susceptible to P. shumwayae and its associated toxin than C. variegatus and implicate differences in the ability these species to osmoregulate as a contributing factor for this phenomenon. Salinity stress enhanced susceptibility of O. niloticus to PfTx and thus improved the sensitivity of the bioassay. The observation that salinity stress enhances toxicity to O. niloticus provides additional information regarding the mechanism of PfTx toxicity although the conditions utilized are not representative of the natural habitat of these freshwater fish. 相似文献
999.
Soil water availability for plants as quantified by conventional available water, least limiting water range and integral water capacity 总被引:1,自引:0,他引:1
Hossein Asgarzadeh Mohammad Reza Mosaddeghi Ali Akbar Mahboubi Akram Nosrati Anthony Roger Dexter 《Plant and Soil》2010,335(1-2):229-244
There are different approaches to define the soil available water (SAW) for plants. The objectives of this study are to evaluate the SAW values of 12 arable soils from Hamadan province (western Iran) calculated by plant available water (PAW), least limiting water range (LLWR) and integral water capacity (IWC) approaches and to explore their relations with Dexter’s index of soil physical quality (i.e., S-value). Soil water retention and mechanical resistance were determined on the intact samples which were taken from the 5–10 cm layer. For calculation of LLWR and IWC, the van Genuchten-Mualem model was fitted to the observed soil water retention data. Two matric suctions (h) of 100 and 330 cm were used for the field capacity (FC). There were significant differences (P?<?0.01) between the SAW values calculated by PAW100, PAW330, LLWR100, LLWR330 and IWC. The highest (i.e., 0.210 cm3 cm?3) and the lowest (i.e., 0.129 cm3 cm?3) means of SAW were calculated for the IWC and LLWR330, respectively. The upper limit of LLWR330 for all of the soils was h of 330 cm, and that of LLWR100 (except for one soil that was air-filled porosity of 0.1 cm3 cm?3) was h of 100 cm. The lower limit of LLWR330 and LLWR100 for five soils was h of 15,000 cm and for seven soils was mechanical resistance of 2 MPa. The IWC values were smaller than those of LLWR100 for two soils, equal to those of LLWR100 for three soils and greater than those of LLWR100 for the rest. There is, therefore, a tendency to predict more SAW using the IWC approach than with the LLWR approach. This is due to the chosen critical soil limits and gradual changes of soil limitations vs. water content in the IWC calculation procedure. Significant relationships of SAW with bulk density or relative bulk density were found but not with the clay and organic matter contents. Linear relations between IWC and LLWR100 or LLWR330 were found as: IWC?=??0.0514 + 1.4438LLWR100, R 2?=?0.83; and IWC?=??0.0405 + 2.0465LLWR330, R 2?=?0.84, respectively (both significant at P?<?0.01). Significant relationships were obtained between the SAW values and S indicating the suitability of the index S to explain the availability of soil water for plants even when complicated approaches like IWC are considered. Overall, the results demonstrate the importance of the choice of the approach to be used and its critical limits in the estimation of the soil available water to plants. 相似文献
1000.