<i>In‐Cell</i><scp>NMR</scp>

Mateos, Borja; Maffei, Mariano; Pons, Miquel

doi:10.1002/9780470015902.a0027154

Cited by 3 publications

(5 citation statements)

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“…[15] For more details on in-cell NMR, we recommend other reviews that focus on this topic. [7,[16][17][18] When investigating membrane receptor interactions on the cell surface the details of the extracellular matrix has also to be considered, as it contains a significant number of polysaccharides (both in free form or as a part of protein glycosylation sites), important for several molecular recognition events, and secondary metabolites that are relevant for intercellular communication. [19] Therefore, the ECM not only supports the mechanical stability of the cells but also is essential for outsidein or inside-out signalling mechanism.…”

Section: The Cellular Environmentmentioning

confidence: 99%

“…Crowding and confinement have important consequences for protein stability and reactivity, as proteins can interact with cellular milieu molecules, either by excluded volume of preferential interactions, [14] and are altered by post‐translational modifications (PTMs) [15] . For more details on in‐cell NMR, we recommend other reviews that focus on this topic [7,16–18] …”

Section: Nmr In Biological Environmentsmentioning

confidence: 99%

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On‐Cell NMR Contributions to Membrane Receptor Binding Characterization

2021

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NMR spectroscopy has matured into a powerful tool to characterize interactions between biological molecules at atomic resolution, most importantly even under near to native (physiological) conditions. The field of in-cell NMR aims to study proteins and nucleic acids inside living cells. However, cells interrogate their environment and are continuously modulated by external stimuli. Cell signaling processes are often initialized by membrane receptors on the cell surface; therefore, characterizing their interactions at atomic resolution by NMR, hereafter referred as on-cell NMR, can provide valuable mechanistic information. This review aims to summarize recent on-cell NMR tools that give information about the binding site and the affinity of membrane receptors to their ligands together with potential applications to in vivo drug screening systems.

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Section: The Cellular Environmentmentioning

confidence: 99%

Section: Nmr In Biological Environmentsmentioning

confidence: 99%

On‐Cell NMR Contributions to Membrane Receptor Binding Characterization

2021

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“…In this study, we introduce methylcellulose (MeCe) hydrogels for NMR studies of extracellular proteins interacting with receptors exposed on the membranes of living cells (ex cell NMR). Although here we focus on protein interactions between cellular receptors and proteins from the extracellular space our findings are also highly relevant for ongoing in‐cell NMR experiments under physiological conditions, for which cell integrity and viability are essential, as well as metabolomic studies …”

Section: Figurementioning

confidence: 99%

“…Figure adapted with permission from ref. , Copyright John Wiley & Sons, Ltd., 2018. B) Normalized STD amplifications from the MeCe (compared with a condition in buffer alone) (top) or the HEK293T cells (compared with a condition in MeCe alone) (bottom) of the wild‐type (left) and the R138A mutant (right).…”

Section: Figurementioning

confidence: 99%

NMR Characterization of Surface Receptor Protein Interactions in Live Cells Using Methylcellulose Hydrogels

et al. 2020

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Interactions of transmembrane receptors with their extracellular ligands are essential for cellular communication and signaling and are therefore a major focus in drug discovery programs. The transition from in vitro to live cell interaction studies, however, is typically a bottleneck in many drug discovery projects due to the challenge of obtaining atomic‐resolution information under near‐physiological conditions. Although NMR spectroscopy is ideally suited to overcome this limitation, several experimental impairments are still present. Herein, we propose the use of methylcellulose hydrogels to study extracellular proteins and their interactions with plasma membrane receptors. This approach reduces cell sedimentation, prevents the internalization of membrane receptors, and increases cell survival, while retaining the free tumbling of extracellular proteins.

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“…In this study, we introduce methylcellulose (MeCe) hydrogels for NMR studies of extracellular proteins interacting with receptors exposed on the membranes of living cells (ex cell NMR). Although here we focus on protein interactions between cellular receptors and proteins from the extracellular space our findings are also highly relevant for ongoing in-cell NMR experiments [23][24][25][26] under physiological conditions, [27,28] for which cell integrity and viability are essential, as well as metabolomic studies. [45] MeCe is particularly favorable for these studies as it is a hydrophilic polymer derived from cellulose and has a wide range of cell biology applications, such as 3D cell cultures.…”

mentioning

confidence: 99%