| 11. |
- Abelein, A, et al.
(author)
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High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain
- 2020
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In: Scientific reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 10:1, s. 235-
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Journal article (peer-reviewed)abstract
- During storage in the silk gland, the N-terminal domain (NT) of spider silk proteins (spidroins) keeps the aggregation-prone repetitive region in solution at extreme concentrations. We observe that NTs from different spidroins have co-evolved with their respective repeat region, and now use an NT that is distantly related to previously used NTs, for efficient recombinant production of the amyloid-β peptide (Aβ) implicated in Alzheimer’s disease. A designed variant of NT from Nephila clavipes flagelliform spidroin, which in nature allows production and storage of β-hairpin repeat segments, gives exceptionally high yields of different human Aβ variants as a solubility tag. This tool enables efficient production of target peptides also in minimal medium and gives up to 10 times more isotope-labeled monomeric Aβ peptides per liter bacterial culture than previously reported.
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| 12. |
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| 13. |
- Askarieh, Glareh, et al.
(author)
-
Self-assembly of spider silk proteins is controlled by a pH-sensitive relay
- 2010
-
In: Nature. - : Springer Nature. - 0028-0836 .- 1476-4687. ; 465:7295, s. 236-8
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Journal article (peer-reviewed)abstract
- Nature's high-performance polymer, spider silk, consists of specific proteins, spidroins, with repetitive segments flanked by conserved non-repetitive domains. Spidroins are stored as a highly concentrated fluid dope. On silk formation, intermolecular interactions between repeat regions are established that provide strength and elasticity. How spiders manage to avoid premature spidroin aggregation before self-assembly is not yet established. A pH drop to 6.3 along the spider's spinning apparatus, altered salt composition and shear forces are believed to trigger the conversion to solid silk, but no molecular details are known. Miniature spidroins consisting of a few repetitive spidroin segments capped by the carboxy-terminal domain form metre-long silk-like fibres irrespective of pH. We discovered that incorporation of the amino-terminal domain of major ampullate spidroin 1 from the dragline of the nursery web spider Euprosthenops australis (NT) into mini-spidroins enables immediate, charge-dependent self-assembly at pH values around 6.3, but delays aggregation above pH 7. The X-ray structure of NT, determined to 1.7 A resolution, shows a homodimer of dipolar, antiparallel five-helix bundle subunits that lack homologues. The overall dimeric structure and observed charge distribution of NT is expected to be conserved through spider evolution and in all types of spidroins. Our results indicate a relay-like mechanism through which the N-terminal domain regulates spidroin assembly by inhibiting precocious aggregation during storage, and accelerating and directing self-assembly as the pH is lowered along the spider's silk extrusion duct.
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| 14. |
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| 15. |
- Davis, B, et al.
(author)
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Voices of chemical biology
- 2021
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In: Nature chemical biology. - : Springer Science and Business Media LLC. - 1552-4469 .- 1552-4450. ; 17:1, s. 1-4
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Journal article (peer-reviewed)
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| 16. |
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| 17. |
- Gonska, Nathalie, et al.
(author)
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Structure-Function Relationship of Artificial Spider Silk Fibers Produced by Straining Flow Spinning
- 2020
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In: Biomacromolecules. - : American Chemical Society (ACS). - 1525-7797 .- 1526-4602. ; 21:6, s. 2116-2124
-
Journal article (peer-reviewed)abstract
- The production of large quantities of artificial spider silk fibers that match the mechanical properties of the native material has turned out to be challenging. Recent advancements in the field make biomimetic spinning approaches an attractive way forward since they allow the spider silk proteins to assemble into the secondary, tertiary, and quaternary structures that are characteristic of the native silk fiber. Straining flow spinning (SFS) is a newly developed and versatile method that allows production under a wide range of processing conditions. Here, we use a recombinant spider silk protein that shows unprecedented water solubility and that is capable of native-like assembly, and we spin it into fibers by the SFS technique. We show that fibers may be spun using different hydrodynamical and chemical conditions and conclude that these spinning conditions affect fiber mechanics. In particular, it was found that the addition of acetonitrile and polyethylene glycol to the collection bath results in fibers with increased beta-sheet content and improved mechanical properties.
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| 18. |
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| 19. |
- Kramer, Lilith, et al.
(author)
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New paths for modelling freshwater nature futures
- 2023
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In: Sustainability Science. - 1862-4065 .- 1862-4057.
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Journal article (peer-reviewed)abstract
- Freshwater ecosystems are exceptionally rich in biodiversity and provide essential benefits to people. Yet they are disproportionately threatened compared to terrestrial and marine systems and remain underrepresented in the scenarios and models used for global environmental assessments. The Nature Futures Framework (NFF) has recently been proposed to advance the contribution of scenarios and models for environmental assessments. This framework places the diverse relationships between people and nature at its core, identifying three value perspectives as points of departure: Nature for Nature, Nature for Society, and Nature as Culture. We explore how the NFF may be implemented for improved assessment of freshwater ecosystems. First, we outline how the NFF and its main value perspectives can be translated to freshwater systems and explore what desirable freshwater futures would look like from each of the above perspectives. Second, we review scenario strategies and current models to examine how freshwater modelling can be linked to the NFF in terms of its aims and outcomes. In doing so, we also identify which aspects of the NFF framework are not yet captured in current freshwater models and suggest possible ways to bridge them. Our analysis provides future directions for a more holistic freshwater model and scenario development and demonstrates how society can benefit from freshwater modelling efforts that are integrated with the value-perspectives of the NFF.
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| 20. |
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