ORCID Profile
0000-0002-4694-4299
Current Organisations
IT University of Copenhagen
,
Københavns Universitet
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Publisher: Cold Spring Harbor Laboratory
Date: 21-08-2021
DOI: 10.1101/2021.08.20.457080
Abstract: The CorA family of proteins regulates the homeostasis of alent metal ions in many bacteria, archaea, and eukaryotic mitochondria, making it an important target in the investigation of the mechanisms of transport and its functional regulation. Although numerous structures of open and closed channels are now available for the CorA family, the mechanism of the transport regulation remains elusive. Here, we investigated the conformational distribution and associated dynamic behaviour of the pentameric Mg 2+ channel CorA at room temperature using small-angle neutron scattering (SANS) in combination with molecular dynamics (MD) simulations and solid-state nuclear magnetic resonance spectroscopy (NMR). We find that neither the Mg 2+ -bound closed structure nor the Mg 2+ -free open forms are sufficient to explain the average conformation of CorA. Our data support the presence of conformational equilibria between multiple states, and we further find a variation in the behaviour of the backbone dynamics with and without Mg 2+ . We propose that CorA must be in a dynamic equilibrium between different non-conducting states, both symmetric and asymmetric, regardless of bound Mg 2+ but that conducting states become more populated in Mg 2+ -free conditions. These properties are regulated by backbone dynamics and are key to understanding the functional regulation of CorA.
Publisher: EMBO
Date: 30-04-2020
Publisher: Wiley
Date: 30-12-2017
DOI: 10.1111/FEBS.14345
Abstract: A novel and generally applicable method for determining structures of membrane proteins in solution via small-angle neutron scattering (SANS) is presented. Common detergents for solubilizing membrane proteins were synthesized in isotope-substituted versions for utilizing the intrinsic neutron scattering length difference between hydrogen and deuterium. In idual hydrogen/deuterium levels of the detergent head and tail groups were achieved such that the formed micelles became effectively invisible in heavy water (D
Publisher: American Chemical Society (ACS)
Date: 05-08-2021
DOI: 10.1021/ACS.MOLPHARMACEUT.1C00164
Abstract: Two different insulin analogues, insulin degludec and lithocholyl insulin, were studied by small-angle X-ray scattering with respect to their self-assembly and interactions in solution at different concentrations of insulin and salt, NaCl. Very different behavior was observed for the two. Insulin degludec, linked to a hexadecanedioic acid, consistently formed di-hexamers, without any further oligomeric growth upon screening of electrostatic repulsions, indicating a stable di-hexamer unit without further oligomerization, as expected in the presence of phenol. The other insulin analogue, linked to the sterol lithocholic acid, formed
Publisher: Public Library of Science (PLoS)
Date: 07-2015
Publisher: Wiley
Date: 12-02-2019
DOI: 10.1111/FEBS.14766
Abstract: 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 MSP 1E3D1, we designed an optimized membrane scaffold protein (His‐ls MSP 1E3D1) 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 (cs MSP 1E3D1) 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 s les. First, we show that circularized and noncircularized (ls MSP 1E3D1) nanodiscs are structurally similar and have the expected nanodisc structure. Second, we show that ls MSP 1E3D1 nanodiscs are more stable compared to the template MSP 1E3D1 nanodiscs as an effect of the extra negative charges and that cs MSP 1E3D1 nanodiscs have further improved stability as an effect of circularization. Finally, we show that a membrane protein can be efficiently incorporated in cs MSP 1E3D1 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.
Publisher: eLife Sciences Publications, Ltd
Date: 07-02-2022
DOI: 10.7554/ELIFE.71887
Abstract: The CorA family of proteins regulates the homeostasis of alent metal ions in many bacteria, archaea, and eukaryotic mitochondria, making it an important target in the investigation of the mechanisms of transport and its functional regulation. Although numerous structures of open and closed channels are now available for the CorA family, the mechanism of the transport regulation remains elusive. Here, we investigated the conformational distribution and associated dynamic behaviour of the pentameric Mg 2+ channel CorA at room temperature using small-angle neutron scattering (SANS) in combination with molecular dynamics (MD) simulations and solid-state nuclear magnetic resonance spectroscopy (NMR). We find that neither the Mg 2+ -bound closed structure nor the Mg 2+ -free open forms are sufficient to explain the average conformation of CorA. Our data support the presence of conformational equilibria between multiple states, and we further find a variation in the behaviour of the backbone dynamics with and without Mg 2+ . We propose that CorA must be in a dynamic equilibrium between different non-conducting states, both symmetric and asymmetric, regardless of bound Mg 2+ but that conducting states become more populated in Mg 2+ -free conditions. These properties are regulated by backbone dynamics and are key to understanding the functional regulation of CorA.
Publisher: eLife Sciences Publications, Ltd
Date: 03-02-2022
No related grants have been discovered for Lise Arleth.