One of the mechanisms by which toxic metal ions interfere with cellular functions is ionic mimicry, where they bind to protein sites in lieu of native metals Ca ^(2+) and Zn ^(2+) . The influence of crowded intracellular environments on these interactions is not well understood. Here, we demonstrate the application of in-cell and lysate NMR spectroscopy to obtain atomic-level information on how a potent environmental toxin cadmium interacts with its protein targets. The experiments, conducted in...
[NMR paper] Diffusion NMR and Rheology of a Model Polymer in Bacterial Cell Lysate Crowders
Diffusion NMR and Rheology of a Model Polymer in Bacterial Cell Lysate Crowders
The intracellular milieu is crowded and heterogeneous, and this can have profound consequences for biomolecule motions and biochemical kinetics. Macromolecular crowding has been traditionally studied in artificial crowders like Ficoll and dextran or globular proteins such as bovine serum albumin. It is, however, not clear if the effects of artificial crowders on such phenomena are the same as the crowding that is experienced in a heterogeneous biological environment. Bacterial cells, for...
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05-22-2023 11:09 PM
[NMR paper] Fluorine NMR spectroscopy enables to quantify the affinity between DNA and proteins in cell lysate
Fluorine NMR spectroscopy enables to quantify the affinity between DNA and proteins in cell lysate
The determination of the binding affinity quantifying the interaction between proteins and nucleic acids is of crucial interest in biological and chemical research. Here, we have made use of sitespecific fluorine labeling of the cold shock protein from Bacillus subtiliis , Bs CspB, enabling to directly monitor the interaction with single stranded DNA molecules in cell lysate. High-resolution 19 F NMR spectroscopy has been applied to exclusively report on resonance signals arising from the......
[NMR paper] All atom insights into the impact of crowded environments on protein stability by NMR spectroscopy.
All atom insights into the impact of crowded environments on protein stability by NMR spectroscopy.
Related Articles All atom insights into the impact of crowded environments on protein stability by NMR spectroscopy.
Nat Commun. 2020 Nov 13;11(1):5760
Authors: Köhn B, Kovermann M
Abstract
The high density of macromolecules affecting proteins due to volume exclusion has been discussed in theory but numerous in vivo experiments cannot be sufficiently understood taking only pure entropic stabilization into account. Here, we show...
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11-17-2020 07:53 AM
[NMR paper] Insights into protein stability in cell lysate by 19F NMR spectroscopy.
Insights into protein stability in cell lysate by 19F NMR spectroscopy.
Related Articles Insights into protein stability in cell lysate by 19F NMR spectroscopy.
Chembiochem. 2020 Aug 12;:
Authors: Welte H, Kovermann M
Abstract
In living organisms, protein folding and function take place in an inhomogeneous, highly crowded environment possessing a concentration of diverse macromolecules of up to 400*g/L. It has been shown that the intracellular environment has a pronounced effect on the stability, dynamics and function of the...
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08-15-2020 05:51 AM
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy.
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy.
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy.
Biochemistry. 2011 Sep 26;
Authors: Wang Q, Zhuravleva A, Gierasch LM
Abstract
Biology relies on functional interplay of proteins in the crowded and heterogeneous environment inside cells, and functional protein interactions are often weak and transient. Thus, methods are needed that preserve these interactions...
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09-30-2011 06:00 AM
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy.
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy.
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy.
Biochemistry. 2011 Sep 26;
Authors: Wang Q, Zhuravleva A, Gierasch LM
Abstract
Biology relies on functional interplay of proteins in the crowded and heterogeneous environment inside cells, and functional protein interactions are often weak and transient. Thus, methods are needed that preserve these...
nmrlearner
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09-30-2011 05:59 AM
Calcium binding environments probed by (43)Ca NMR spectroscopy.
Calcium binding environments probed by (43)Ca NMR spectroscopy.
Calcium binding environments probed by (43)Ca NMR spectroscopy.
Dalton Trans. 2010 Oct 7;39(37):8593-602
Authors: Bryce DL
Calcium is an important component of materials, metalloproteins, minerals, glasses, and small inorganic and organic complexes. However, NMR spectroscopy of the quadrupolar (43)Ca nuclide remains difficult primarily due to its low natural abundance and low resonance frequency. In this Perspective, experimental challenges and recent successes in the field are...
Protein-Cadmium Interactions in Crowded Biomolecular Environments Probed by In-cell and Lysate NMR Spectroscopy
Linear Mode
