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  • 1
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    Unbekannt
    The American Society for Biochemistry and Molecular Biology (ASBMB)
    Publikationsdatum: 2016-02-20
    Beschreibung: Prostaglandin endoperoxide H synthases (PGHSs), also called cyclooxygenases (COXs), convert arachidonic acid (AA) to PGH2. PGHS-1 and PGHS-2 are conformational heterodimers, each composed of an (Eallo) and a catalytic (Ecat) monomer. Previous studies suggested that the binding to Eallo of saturated or monounsaturated fatty acids (FAs) that are not COX substrates differentially regulate PGHS-1 versus PGHS-2. Here, we substantiate and expand this concept to include polyunsaturated FAs known to modulate COX activities. Non-substrate FAs like palmitic acid bind Eallo of PGHSs stimulating human (hu) PGHS-2 but inhibiting huPGHS-1. We find the maximal effects of non-substrate FAs on both huPGHSs occurring at the same physiologically relevant FA/AA ratio of ∼20. This inverse allosteric regulation likely underlies the ability of PGHS-2 to operate at low AA concentrations, when PGHS-1 is effectively latent. Unlike FAs tested previously, we observe that C-22 FAs, including ω-3 fish oil FAs, have higher affinities for Ecat than Eallo subunits of PGHSs. Curiously, C-20 ω-3 eicosapentaenoate preferentially binds Ecat of huPGHS-1 but Eallo of huPGHS-2. PGE2 production decreases 50% when fish oil consumption produces tissue EPA/AA ratios of ≥0.2. However, 50% inhibition of huPGHS-1 itself is only seen with ω-3 FA/AA ratios of ≥5.0. This suggests that fish oil-enriched diets disfavor AA oxygenation by altering the composition of the FA pool in which PGHS-1 functions. The distinctive binding specificities of PGHS subunits permit different combinations of non-esterified FAs, which can be manipulated dietarily, to regulate AA binding to Eallo and/or Ecat thereby controlling COX activities.
    Print ISSN: 0021-9258
    Digitale ISSN: 1083-351X
    Thema: Biologie , Chemie und Pharmazie
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 2
    facet.materialart.
    Unbekannt
    The American Society for Biochemistry and Molecular Biology (ASBMB)
    Publikationsdatum: 2013-10-05
    Beschreibung: Prostaglandin endoperoxide H synthase-2 (PGHS-2), also known as cyclooxygenase-2 (COX-2), is a sequence homodimer. However, the enzyme exhibits half-site heme and inhibitor binding and functions as a conformational heterodimer having a catalytic subunit (Ecat) with heme bound and an allosteric subunit (Eallo) lacking heme. Some recombinant heterodimers composed of a COX-deficient mutant subunit and a native subunit (i.e. Mutant/Native PGHS-2) have COX activities similar to native PGHS-2. This suggests that the presence of heme plus substrate leads to the subunits becoming lodged in a semi-stable Eallo-mutant/Ecat-Native∼heme form during catalysis. We examined this concept using human PGHS-2 dimers composed of combinations of Y385F, R120Q, R120A, and S530A mutant or native subunits. With some heterodimers (e.g. Y385F/Native PGHS-2), heme binds with significantly higher affinity to the native subunit. This correlates with near native COX activity for the heterodimer. With other heterodimers (e.g. S530A/Native PGHS-2), heme binds with similar affinities to both subunits, and the COX activity approximates that expected for an enzyme in which each monomer contributes equally to the net COX activity. With or without heme, aspirin acetylates one-half of the subunits of the native PGHS-2 dimer, the Ecat subunits. Subunits having an S530A mutation are refractory to acetylation. Curiously, aspirin acetylates only one-quarter of the monomers of S530A/Native PGHS-2 with or without heme. This implies that there are comparable amounts of two noninterchangeable species of apoenzymes, Eallo-S530A/Ecat-Native and Eallo-Native/Ecat-S530A. These results suggest that native PGHS-2 assumes a reasonably stable, asymmetric Eallo/Ecat form during its folding and processing.
    Print ISSN: 0021-9258
    Digitale ISSN: 1083-351X
    Thema: Biologie , Chemie und Pharmazie
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 3
    facet.materialart.
    Unbekannt
    The American Society for Biochemistry and Molecular Biology (ASBMB)
    Publikationsdatum: 2012-10-13
    Beschreibung: In the early 1900s, dietary fat was viewed simply as a source of calories, interchangeable with carbohydrates, but in 1929 and 1930, a husband-and-wife team published two papers in the Journal of Biological Chemistry that turned the notion on its head. Through meticulous analyses of rats fed special diets, George and Mildred Burr discovered that fatty acids were critical to health. If fatty acids were missing in the diet, a deficiency syndrome ensued that often led to death. The Burrs identified linoleic acid as an essential fatty acid and coined the phrase “essential fatty acids.” jbc;287/42/35439/FU1F1FU1 George O. Burr in 1980. This photograph was provided courtesy of the Eskind Biomedical Library, Vanderbilt University Medical Center. The work by the Burrs “showed that fats are not there solely as calories to support growth but that they are important for proper physiology,” explains Norman Salem, Jr., of DSM Nutritional Products, a company that makes bulk vitamins, lipids, carotenoids, and other nutrition products. The two papers heralded “the beginning of a modern paradigm in nutritional biochemistry.” The field of nutritional fatty acid research has exploded since the work by the Burrs and now affects our daily lives. Food manufacturers add fatty acid supplements, such as the omega-3 fatty acids eicosapentaenoic and docosahexaenoic acids (more popularly known as EPA and DHA), to processed foods, and government agencies work to establish guidelines on which fats should be incorporated into healthy diets. In a speech he gave in 1980 at the Golden Jubilee International Congress...
    Print ISSN: 0021-9258
    Digitale ISSN: 1083-351X
    Thema: Biologie , Chemie und Pharmazie
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 4
    facet.materialart.
    Unbekannt
    The American Society for Biochemistry and Molecular Biology (ASBMB)
    Publikationsdatum: 2015-02-28
    Beschreibung: Cyclooxygenases (COXs) catalyze the committed step in prostaglandin (PG) biosynthesis. COX-1 is constitutively expressed and stable, whereas COX-2 is inducible and short lived. COX-2 is degraded via endoplasmic reticulum (ER)-associated degradation (ERAD) following post-translational glycosylation of Asn-594. COX-1 and COX-2 are found in abundance on the luminal surfaces of the ER and inner membrane of the nuclear envelope. Using confocal immunocytofluorescence, we detected both COX-2 and microsomal PGE synthase-1 (mPGES-1) but not COX-1 in the Golgi apparatus. Inhibition of trafficking between the ER and Golgi retarded COX-2 ERAD. COX-2 has a C-terminal STEL sequence, which is an inefficient ER retention signal. Substituting this sequence with KDEL, a robust ER retention signal, concentrated COX-2 in the ER where it was stable and slowly glycosylated on Asn-594. Native COX-2 and a recombinant COX-2 having a Golgi targeting signal but not native COX-1 exhibited efficient catalytic coupling to mPGES-1. We conclude that N-glycosylation of Asn-594 of COX-2 occurs in the ER, leading to anterograde movement of COX-2 to the Golgi where the Asn-594-linked glycan is trimmed prior to retrograde COX-2 transport to the ER for ERAD. Having an inefficient ER retention signal leads to sluggish Golgi to ER transit of COX-2. This permits significant Golgi residence time during which COX-2 can function catalytically. Cytosolic phospholipase A2α, which mobilizes arachidonic acid for PG synthesis, preferentially translocates to the Golgi in response to physiologic Ca2+ mobilization. We propose that cytosolic phospholipase A2α, COX-2, and mPGES-1 in the Golgi comprise a dedicated system for COX-2-dependent PGE2 biosynthesis.
    Print ISSN: 0021-9258
    Digitale ISSN: 1083-351X
    Thema: Biologie , Chemie und Pharmazie
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 5
    ISSN: 1471-4159
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Notizen: Abstract: Electrophysiological studies of photoreceptors from the horseshoe crab Limulus polyphemus continue to provide fundamental new knowledge of the photoresponse in invertebrates. Therefore, it is of particular interest to characterize the molecular components of the photoresponse in this system. Here we describe an arrestin cloned from a cDNA library constructed using poly(A)+ RNA isolated from Limulus lateral eyes. The protein, deduced from the arrestin cDNA, is most similar to arrestin from locust antennae (56% identity) and Drosophila phosrestin I (53% identity). Limulus arrestin was expressed in a heterologous system, and its properties were compared with those of a 46-kDa light-regulated phosprotein (pp46A) in Limulus photoreceptors described in previous studies from this laboratory. Arrestin and pp46A (a) have the same apparent molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, (b) have an isoelectric point in the basic pH range, (c) require calmodulin and elevated Ca2+ levels for phosphorylation, (d) are immunoreactive with monoclonal antibody C10C10 directed against a sequence in bovine arrestin (S-antigen) that is perfectly conserved in the deduced arrestin protein, and (e) are associated with photoreceptors. We conclude that the arrestin described here and pp46A are the same protein. The results of this and previous studies show that in Limulus photoreceptors, light regulates the phosphorylation of arrestin in complex ways.
    Materialart: Digitale Medien
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 6
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Science Ltd
    Global change biology 6 (2000), S. 0 
    ISSN: 1365-2486
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Biologie , Energietechnik , Geographie
    Notizen: Changes in soil organic carbon (SOC) in agricultural soils influence soil quality and greenhouse gas concentrations in the atmosphere. Land use, management practices, soil characteristics, and climate influence such changes. Using the Century model we estimated the rate of SOC change in agricultural soils of Canada for the period 1970 to 2010. This estimation was based on the estimated SOC change for 15% of the 1250 agriculturally designated soil landscape of Canada (SLC) polygons. Simulations were carried out for two to five crop rotations and for conventional and no-tillage. The results indicate that the agricultural soils in Canada, whose SOC are currently very close to equilibrium, will stop being a net source of CO2 and will become a sink by the year 2000. Rates of carbon change for the years 1970, 1990, and 2010 were estimated to be −67, − 39, and 11 kgC ha−1. The rate of decline in the carbon content of agricultural soils in Canada has slowed considerably in the 1990s as a result of an increase in the adoption of no-tillage management, a reduction in the use of summer fallowing, and an increase in fertilizer application. We estimate that the proportion of agricultural land storing SOC will have increased from 17% in 1990 to 53% by the year 2000.
    Materialart: Digitale Medien
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 7
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Science Ltd
    Clinical & experimental allergy 34 (2004), S. 0 
    ISSN: 1365-2222
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Notizen: Background Cat allergy is unique among allergy to mammals in that the major allergen Fel d 1 is a uteroglobin-like protein and not a lipocalin. The biochemical spectrum of the cat allergens is thus uncertain, particularly with regard to the role that a cat lipocalin protein may play in sensitization to cats in allergic individuals.Objective To analyse cDNA encoding a lipocalin allergen and the corresponding recombinant allergen at both the molecular and immunological levels.Methods A submandibular salivary gland cDNA expression library was constructed and screened for clones producing IgE-binding polypeptides. cDNA encoding a lipocalin allergen and its corresponding recombinant allergen were analysed.Results An IgE binding molecule with high sequence identity to the boar salivary lipocalin and the horse lipocalin Equ c 1 allergen was isolated and designated, Fel d 4. Serum from 62.96% of cat-allergic subjects examined had measurable IgE antibody to Fel d 4 but typically at low levels. Despite this in 47% of sera the anti-Fel d 4 IgE titres were higher than the anti-Fel d 1 titres. IgE binding to the lipocalin allergen could be blocked by an allergen extract from cow and to a lesser degree by extracts from horse and dog.Conclusion Fel d 4 is a lipocalin allergen produced by the cat, which binds IgE at relatively high frequency in cat-sensitive individuals. The allergen provides not only a means for investigating differences in the immune response to lipocalin allergens from that found for other mammalian species but also an important reagent for the diagnosis of cat allergy.
    Materialart: Digitale Medien
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 8
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Science Ltd
    Clinical & experimental allergy 33 (2003), S. 0 
    ISSN: 1365-2222
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Materialart: Digitale Medien
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 9
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Science Ltd
    Clinical & experimental allergy 32 (2002), S. 0 
    ISSN: 1365-2222
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Materialart: Digitale Medien
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 10
    Digitale Medien
    Digitale Medien
    Oxford BSL : Blackwell Science Ltd
    Clinical & experimental allergy 29 (1999), S. 0 
    ISSN: 1365-2222
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Materialart: Digitale Medien
    Standort Signatur Einschränkungen Verfügbarkeit
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