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  • 1
    In: Science, American Association for the Advancement of Science (AAAS), Vol. 378, No. 6616 ( 2022-10-14), p. 186-192
    Abstract: Single-molecule peptide sequences are determined by N-terminal amino acid recognizers.
    Type of Medium: Online Resource
    ISSN: 0036-8075 , 1095-9203
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    Language: English
    Publisher: American Association for the Advancement of Science (AAAS)
    Publication Date: 2022
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  • 2
    In: Proceedings of the National Academy of Sciences, Proceedings of the National Academy of Sciences, Vol. 114, No. 41 ( 2017-10-10)
    Abstract: Circadian clocks play an important role in lipid homeostasis, with impact on various metabolic diseases. Due to the central role of skeletal muscle in whole-body metabolism, we aimed at studying muscle lipid profiles in a temporal manner. Moreover, it has not been shown whether lipid oscillations in peripheral tissues are driven by diurnal cycles of rest–activity and food intake or are able to persist in vitro in a cell-autonomous manner. To address this, we investigated lipid profiles over 24 h in human skeletal muscle in vivo and in primary human myotubes cultured in vitro. Glycerolipids, glycerophospholipids, and sphingolipids exhibited diurnal oscillations, suggesting a widespread circadian impact on muscle lipid metabolism. Notably, peak levels of lipid accumulation were in phase coherence with core clock gene expression in vivo and in vitro. The percentage of oscillating lipid metabolites was comparable between muscle tissue and cultured myotubes, and temporal lipid profiles correlated with transcript profiles of genes implicated in their biosynthesis. Lipids enriched in the outer leaflet of the plasma membrane oscillated in a highly coordinated manner in vivo and in vitro. Lipid metabolite oscillations were strongly attenuated upon siRNA-mediated clock disruption in human primary myotubes. Taken together, our data suggest an essential role for endogenous cell-autonomous human skeletal muscle oscillators in regulating lipid metabolism independent of external synchronizers, such as physical activity or food intake.
    Type of Medium: Online Resource
    ISSN: 0027-8424 , 1091-6490
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    Language: English
    Publisher: Proceedings of the National Academy of Sciences
    Publication Date: 2017
    detail.hit.zdb_id: 209104-5
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  • 3
    Online Resource
    Online Resource
    American Association for the Advancement of Science (AAAS) ; 1997
    In:  Science Vol. 275, No. 5299 ( 1997-01-24), p. 523-527
    In: Science, American Association for the Advancement of Science (AAAS), Vol. 275, No. 5299 ( 1997-01-24), p. 523-527
    Abstract: The nuclear factor κB (NF-κB) transcription factor is responsive to specific cytokines and stress and is often activated in association with cell damage and growth arrest in eukaryotes. NF-κB is a heterodimeric protein, typically composed of 50- and 65-kilodalton subunits of the Rel family, of which RelA(p65) stimulates transcription of diverse genes. Specific cyclin-dependent kinases (CDKs) were found to regulate transcriptional activation by NF-κB through interactions with the coactivator p300. The transcriptional activation domain of RelA(p65) interacted with an amino-terminal region of p300 distinct from a carboxyl-terminal region of p300 required for binding to the cyclin E-Cdk2 complex. The CDK inhibitor p21 or a dominant negative Cdk2, which inhibited p300-associated cyclin E-Cdk2 activity, stimulated κB-dependent gene expression, which was also enhanced by expression of p300 in the presence of p21. The interaction of NF-κB and CDKs through the p300 and CBP coactivators provides a mechanism for the coordination of transcriptional activation with cell cycle progression.
    Type of Medium: Online Resource
    ISSN: 0036-8075 , 1095-9203
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    Language: English
    Publisher: American Association for the Advancement of Science (AAAS)
    Publication Date: 1997
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  • 4
    In: Protein Science, Wiley, Vol. 11, No. 4 ( 2002-04), p. 820-830
    Abstract: The P22 tailspike adhesin is an elongated thermostable trimer resistant to protease digestion and to denaturation in sodium dodecyl sulfate. Monomeric, dimeric, and protrimeric folding and assembly intermediates lack this stability and are thermolabile. In the native trimer, three right‐handed parallel β‐helices (residues 143–540), pack side‐by‐side around the three‐fold axis. After residue 540, these single chain β‐helices terminate and residues 541–567 of the three polypeptide chains wrap around each other to form a three‐stranded interdigitated β‐helix. Three mutants located in this region — G546D, R563Q, and A575T — blocked formation of native tailspike trimers, and accumulated soluble forms of the mutant polypeptide chains within cells. The substitutions R563Q and A575T appeared to prevent stable association of partially folded monomers. G546D, in the interdigitated region of the chain, blocked tailspike folding at the transition from the partially‐folded protrimer to the native trimer. The protrimer‐like species accumulating in the G546D mutant melted out at 42°C and was trypsin and SDS sensitive. The G546D defect was not corrected by introduction of global suppressor mutations, which correct kinetic defects in β‐helix folding. The simplest interpretation of these results is that the very high thermostability (T m = 88°C), protease and detergent resistance of the native tailspike acquired in the protrimer‐to‐trimer transition, depends on the formation of the three‐stranded interdigitated region. This interdigitated β‐helix appears to function as a molecular clamp insuring thermostable subunit association in the native trimer.
    Type of Medium: Online Resource
    ISSN: 0961-8368 , 1469-896X
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    Language: English
    Publisher: Wiley
    Publication Date: 2002
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  • 5
    Online Resource
    Online Resource
    American Physiological Society ; 2021
    In:  Journal of Applied Physiology Vol. 130, No. 5 ( 2021-05-01), p. 1534-1543
    In: Journal of Applied Physiology, American Physiological Society, Vol. 130, No. 5 ( 2021-05-01), p. 1534-1543
    Abstract: Constant routine and forced desynchrony protocols typically remove the effects of behavioral/environmental cues to examine endogenous circadian rhythms, yet this may not reflect rhythms of appetite regulation in the real world. It is therefore important to understand these rhythms within the same subjects under controlled diurnal conditions of light, sleep, and feeding. Ten healthy adults (9 M/1 F, means ±SD: age, 30 ± 10 yr; body mass index, 24.1 ± 2.7 kg·m −2 ) rested supine in the laboratory for 37 h. All data were collected during the final 24 h of this period (i.e., 0800–0800 h). Participants were fed hourly isocaloric liquid meal replacements alongside appetite assessments during waking before a sleep opportunity from 2200 to 0700 h. Hourly blood samples were collected throughout the 24-h period. Dim light melatonin onset occurred at 2318 ± 46 min. A diurnal rhythm in mean plasma unacylated ghrelin concentration was identified ( P = 0.04), with the acrophase occurring shortly after waking (0819), falling to a nadir in the evening with a relative amplitude of 9%. Plasma leptin concentration also exhibited a diurnal rhythm ( P 〈 0.01), with the acrophase occurring shortly after lights-out (0032 h) and the lowest concentrations at midday. The amplitude for this rhythm was 25%. Diurnal rhythms were established in all dimensions of appetite except for sweet preference ( P = 0.29), with both hunger (2103 h) and prospective food consumption (1955 h) reaching their peak in the evening before falling to their nadir shortly after waking. Under controlled diurnal conditions, simultaneous measurement of leptin, unacylated ghrelin, and subjective appetite over a 24-h period revealed rhythmicity in appetite regulation in lean, healthy humans. NEW & NOTEWORTHY Simultaneous assessment of subjective appetite, unacylated ghrelin, and leptin was carried out over a continuous 37-h protocol for the first time under conditions of controlled light, sleep, and feeding in healthy, lean adults. Rhythms were observed in unacylated ghrelin, leptin, and components of subjective appetite, such as hunger, prospective consumption, and fullness. Concurrent measurement of rhythms in these variables is important to fully understand the temporal relationships between components of appetite as well as the influence of diurnal factors such as sleep, light, and feeding.
    Type of Medium: Online Resource
    ISSN: 8750-7587 , 1522-1601
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    Language: English
    Publisher: American Physiological Society
    Publication Date: 2021
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  • 6
    In: Proceedings of the National Academy of Sciences, Proceedings of the National Academy of Sciences, Vol. 110, No. 8 ( 2013-02-19), p. 2804-2809
    Abstract: Multidrug-resistant Staphylococcus aureus infections pose a significant threat to human health. Antibiotic resistance is most commonly propagated by conjugative plasmids like pLW1043, the first vancomycin-resistant S. aureus vector identified in humans. We present the molecular basis for resistance transmission by the nicking enzyme in S. aureus (NES), which is essential for conjugative transfer. NES initiates and terminates the transfer of plasmids that variously confer resistance to a range of drugs, including vancomycin, gentamicin, and mupirocin. The NES N-terminal relaxase–DNA complex crystal structure reveals unique protein–DNA contacts essential in vitro and for conjugation in S. aureus . Using this structural information, we designed a DNA minor groove-targeted polyamide that inhibits NES with low micromolar efficacy. The crystal structure of the 341-residue C-terminal region outlines a unique architecture; in vitro and cell-based studies further establish that it is essential for conjugation and regulates the activity of the N-terminal relaxase. This conclusion is supported by a small-angle X-ray scattering structure of a full-length, 665-residue NES–DNA complex. Together, these data reveal the structural basis for antibiotic multiresistance acquisition by S . aureus and suggest novel strategies for therapeutic intervention.
    Type of Medium: Online Resource
    ISSN: 0027-8424 , 1091-6490
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    Language: English
    Publisher: Proceedings of the National Academy of Sciences
    Publication Date: 2013
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  • 7
    Online Resource
    Online Resource
    Wiley ; 2000
    In:  Protein Science Vol. 9, No. 12 ( 2000-01), p. 2338-2343
    In: Protein Science, Wiley, Vol. 9, No. 12 ( 2000-01), p. 2338-2343
    Abstract: A right‐handed parallel β‐helix of 400 residues in 13 tightly packed coils is a major motif of the chains forming the trimeric P22 tailspike adhesin. The β‐helix domains of three identical subunits are side‐by‐side in the trimer and make predominantly hydrophilic inter‐subunit contacts (Steinbacher S et al., 1994, Science 265 :383‐386). After the 13th coil the three individual β‐helices terminate and the chains wrap around each other to form three interdigitated β‐sheets organized into the walls of a triangular prism. The β‐strands then separate and form antiparallel β‐sheets, but still defining a triangular prism in which each side is a β‐sheet from a different subunit (Seckler R, 1998, J Struct Biol 122 :216–222). The subunit interfaces are buried in the triangular core of the prism, which is densely packed with hydrophobic side chains from the three β‐sheets. Examination of this structure reveals that its packed core maintains the same pattern of interior packing found in the left‐handed β‐helix, a single‐chain structure. This packing is maintained in both the interdigitated parallel region of the prism and the following antiparallel sheet section. This oligomerization motif for the tailspike β‐helices presumably contributes to the very high thermal and detergent stability that is a property of the native tailspike adhesin.
    Type of Medium: Online Resource
    ISSN: 0961-8368 , 1469-896X
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    Language: English
    Publisher: Wiley
    Publication Date: 2000
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  • 8
    Online Resource
    Online Resource
    American Chemical Society (ACS) ; 2016
    In:  Journal of Natural Products Vol. 79, No. 6 ( 2016-06-24), p. 1702-1706
    In: Journal of Natural Products, American Chemical Society (ACS), Vol. 79, No. 6 ( 2016-06-24), p. 1702-1706
    Type of Medium: Online Resource
    ISSN: 0163-3864 , 1520-6025
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    Language: English
    Publisher: American Chemical Society (ACS)
    Publication Date: 2016
    detail.hit.zdb_id: 1491522-4
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    SSG: 15,3
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  • 9
    Online Resource
    Online Resource
    American Chemical Society (ACS) ; 2015
    In:  Journal of Natural Products Vol. 78, No. 8 ( 2015-08-28), p. 2145-2148
    In: Journal of Natural Products, American Chemical Society (ACS), Vol. 78, No. 8 ( 2015-08-28), p. 2145-2148
    Type of Medium: Online Resource
    ISSN: 0163-3864 , 1520-6025
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    Language: English
    Publisher: American Chemical Society (ACS)
    Publication Date: 2015
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    SSG: 15,3
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  • 10
    In: Protein Science, Wiley, Vol. 10, No. 2 ( 2001-02), p. 397-410
    Abstract: The predominantly β‐sheet phage P22 tailspike adhesin contains eight reduced cysteines per 666 residue chain, which are buried and unreactive in the native trimer. In the pathway to the native trimer, both in vivo and in vitro transient interchain disulfide bonds are formed and reduced. This occurs in the protrimer, an intermediate in the formation of the interdigitated β‐sheets of the trimeric tailspike. Each of the eight cysteines was replaced with serine by site‐specific mutagenesis of the cloned P22 tailspike gene and the mutant genes expressed in Escherichia coli . Although the yields of native‐like Cys 〉 Ser proteins varied, sufficient soluble trimeric forms of each of the eight mutants accumulated to permit purification. All eight single Cys 〉 Ser mature proteins maintained the high thermostability of the wild type, as well as the wild‐type biological activity in forming infectious virions. Thus, these cysteine thiols are not required for the stability or activity of the native state. When their in vivo folding and assembly kinetics were examined, six of the mutant substitutions—C267S, C287S, C458S, C613S, and C635S—were significantly impaired at higher temperatures. Four—C290S, C496, C613S, and C635—showed significantly impaired kinetics even at lower temperatures. The in vivo folding of the C613S/C635S double mutant was severely defective independent of temperature. Since the trimeric states of the single Cys 〉 Ser substituted chains were as stable and active as wild type, the impairment of tailspike maturation presumably reflects problems in the in vivo folding or assembly pathways. The formation or reduction of the transient interchain disulfide bonds in the protrimer may be the locus of these kinetic functions.
    Type of Medium: Online Resource
    ISSN: 0961-8368 , 1469-896X
    RVK:
    Language: English
    Publisher: Wiley
    Publication Date: 2001
    detail.hit.zdb_id: 2000025-X
    SSG: 12
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