| People | Locations | Statistics |
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| Naji, M. |
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| Motta, Antonella |
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| Aletan, Dirar |
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| Mohamed, Tarek |
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| Ertürk, Emre |
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| Taccardi, Nicola |
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| Kononenko, Denys |
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| Petrov, R. H. | Madrid |
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| Alshaaer, Mazen | Brussels |
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| Bih, L. |
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| Casati, R. |
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| Muller, Hermance |
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| Kočí, Jan | Prague |
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| Šuljagić, Marija |
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| Kalteremidou, Kalliopi-Artemi | Brussels |
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| Azam, Siraj |
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| Ospanova, Alyiya |
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| Blanpain, Bart |
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| Ali, M. A. |
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| Popa, V. |
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| Rančić, M. |
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| Ollier, Nadège |
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| Azevedo, Nuno Monteiro |
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| Landes, Michael |
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| Rignanese, Gian-Marco |
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Arleth, Lise
University of Copenhagen
in Cooperation with on an Cooperation-Score of 37%
Topics
Publications (15/15 displayed)
- 2023Modeling of flexible membrane-bound biomolecular complexes for solution small-angle scatteringcitations
- 2023Aggregative adherence fimbriae form compact structures as seen by SAXScitations
- 2022Mg2+-dependent conformational equilibria in CorA and an integrated view on transport regulationcitations
- 2022Mg2+-dependent conformational equilibria in CorA and an integrated view on transport regulationcitations
- 2021Mg2+-dependent conformational equilibria in CorA: an integrated view on transport regulation
- 2021The microscopic distribution of hydrophilic polymers in interpenetrating polymer networks (IPNs) of medical grade siliconecitations
- 2020Assessment of structure factors for analysis of small-angle scattering data from desired or undesired aggregatescitations
- 2020Dispersion state of TiO2 pigment particles studied by ultra-small-angle X-ray scattering revealing dependence on dispersant but limited change during drying of paint coatingcitations
- 2019Circularized and solubility-enhanced MSPs facilitate simple and high-yield production of stable nanodiscs for studies of membrane proteins in solutioncitations
- 2016Construction of insulin 18-mer nanoassemblies driven by coordination to Iron(II) and Zinc(II) ions at distinct sitescitations
- 2016Dimeric peptides with three different linkers self-assemble with phospholipids to form peptide nanodiscs that stabilize membrane proteinscitations
- 2015Small-angle scattering determination of the shape and localization of human cytochrome P450 embedded in a phospholipid nanodisc environmentcitations
- 2014Stealth carriers for low-resolution structure determination of membrane proteins in solutioncitations
- 2013Self-assembly of designed coiled coil peptides studied by small-angle X-ray scattering and analytical ultracentrifugationcitations
- 2013WillItFitcitations
Places of action
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article
Circularized and solubility-enhanced MSPs facilitate simple and high-yield production of stable nanodiscs for studies of membrane proteins in solution
Abstract
<p>Recently, an enzymatic reaction was utilized to covalently link the N and C termini of membrane scaffold proteins to produce circularized nanodiscs that were more homogeneous and stable than standard nanodiscs. We continue this development and aim for obtaining high yields of stable and monodisperse nanodiscs for structural studies of membrane proteins by solution small-angle scattering techniques. Based on the template MSP1E3D1, we designed an optimized membrane scaffold protein (His-lsMSP1E3D1) with a sortase recognition motif and high abundance of solubility-enhancing negative charges. With these modifications, we show that high protein expression is maintained and that the circularization reaction is efficient, such that we obtain a high yield of circularized membrane scaffold protein (csMSP1E3D1) and downstream circularized nanodiscs. We characterize the circularized protein and corresponding nanodiscs biophysically by small-angle X-ray scattering, size-exclusion chromatography, circular dichroism spectroscopy, and light scattering and compare to noncircularized samples. First, we show that circularized and noncircularized (lsMSP1E3D1) nanodiscs are structurally similar and have the expected nanodisc structure. Second, we show that lsMSP1E3D1 nanodiscs are more stable compared to the template MSP1E3D1 nanodiscs as an effect of the extra negative charges and that csMSP1E3D1 nanodiscs have further improved stability as an effect of circularization. Finally, we show that a membrane protein can be efficiently incorporated in csMSP1E3D1 nanodiscs. Large-scale production methods for circularized nanodiscs with improved thermal and temporal stability will facilitate better access to the nanodisc technology and enable applications at physiologically relevant temperatures.</p>