Looking into live cells with in-cell NMR spectroscopy

Selenko, Philipp; Wagner, Gerhard

doi:10.1016/j.jsb.2007.04.001

Cited by 132 publications

(98 citation statements)

References 48 publications

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“…With improving isotopic labeling schemes and pulse sequences, whole cell NMR of living cells also promises to soon be applicable to eukaryotic cell types [57]. Similarly, mass spectrometry has been used to probe how the interior of bacterial cells changes protein stability [58], and could be expanded to eukaryotic cells.…”

Section: Leaving the Test-tube Behindmentioning

confidence: 99%

Quinary protein structure and the consequences of crowding in living cells: Leaving the test‐tube behind

2013

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Although the importance of weak protein-protein interactions has been understood since the 1980s, scant attention has been paid to this "quinary structure". The transient nature of quinary structure facilitates dynamic sub-cellular organization through loose grouping of proteins with multiple binding partners. Despite our growing appreciation of the quinary structure paradigm in cell biology, we do not yet understand how the many forces inside the cell--the excluded volume effect, the "stickiness" of the cytoplasm, and hydrodynamic interactions--perturb the weakest functional protein interactions. We discuss the unresolved problem of how the forces in the cell modulate quinary structure, and to what extent the cell has evolved to exert control over the weakest biomolecular interactions. We conclude by highlighting the new experimental and computational tools coming on-line for in vivo studies, which are a critical next step if we are to understand quinary structure in its native environment.

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Section: Leaving the Test-tube Behindmentioning

confidence: 99%

Quinary protein structure and the consequences of crowding in living cells: Leaving the test‐tube behind

2013

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“…I n-cell NMR represents a unique tool for characterizing physiological processes and their alteration in the 'natural' environment, that is, within a live cellular context, with an atomic-level approach [1][2][3][4][5][6][7][8][9] . This level of characterization is necessary to understand the molecular cause of cell malfunction in pathological states and to develop treatments and therapeutic protocols.…”

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confidence: 99%

In-cell NMR reveals potential precursor of toxic species from SOD1 fALS mutants

et al. 2014

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Mutations in the superoxide dismutase 1 (SOD1) gene are related to familial cases of amyotrophic lateral sclerosis (fALS). Here we exploit in-cell NMR to characterize the protein folding and maturation of a series of fALS-linked SOD1 mutants in human cells and to obtain insight into their behaviour in the cellular context, at the molecular level. The effect of various mutations on SOD1 maturation are investigated by changing the availability of metal ions in the cells, and by coexpressing the copper chaperone for SOD1, hCCS. We observe for most of the mutants the occurrence of an unstructured SOD1 species, unable to bind zinc. This species may be a common precursor of potentially toxic oligomeric species, that are associated with fALS. Coexpression of hCCS in the presence of copper restores the correct maturation of the SOD1 mutants and prevents the formation of the unstructured species, confirming that hCCS also acts as a molecular chaperone.

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“…In 2001, they clearly demonstrated that the structure of mercury-detoxification MerA bacterial protein, overexpressed in E. coli, differed from in vitro models . This technique is constantly improved and is a valuable way of providing the most relevant structural information (Maldonado et al 2011;Selenko and Wagner 2007). Recently, the technique was applied to study membrane proteins, a very difficult task due to the resolution and stability limitations induced by the lipid environment.…”

Section: High-resolution Structure In Live Cellsmentioning

confidence: 99%