In-cell NMR spectroscopy is a powerful approach to study protein structure and function in the native cellular environment. It provides precious insights into the folding, maturation, interactions, and ligand binding of important pharmacological targets directly in human cells. However, its widespread application is hampered by the fact that soluble globular proteins often interact with large cellular components, causing severe line broadening in conventional heteronuclear NMR experiments....
[ASAP] Direct Expression of Fluorinated Proteins in Human Cells for 19F In-Cell NMR Spectroscopy
Direct Expression of Fluorinated Proteins in Human Cells for 19F In-Cell NMR Spectroscopy
Lan B. T. Pham, Azzurra Costantino, Letizia Barbieri, Vito Calderone, Enrico Luchinat, and Lucia Banci
https://pubs.acs.org/cms/10.1021/jacs.2c12086/asset/images/medium/ja2c12086_0007.gif
Journal of the American Chemical Society
DOI: 10.1021/jacs.2c12086
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[NMR paper] Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor
Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor
In-cell NMR is a unique approach to observe the structural and dynamic properties of biological macromolecules at atomic resolution directly in living cells. Protein folding, chemical modifications, and conformational changes induced by ligand binding can be observed. Therefore, this method has great potential in the context of drug development. However, the short lifetime of human cells confined in the NMR spectrometer limits the application range of in-cell...
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03-30-2021 08:29 AM
[NMR paper] Observation of nucleic acids inside living human cells by in-cell NMR spectroscopy.
Observation of nucleic acids inside living human cells by in-cell NMR spectroscopy.
Observation of nucleic acids inside living human cells by in-cell NMR spectroscopy.
Biophys Physicobiol. 2020;17:36-41
Authors: Yamaoki Y, Nagata T, Sakamoto T, Katahira M
Abstract
The intracellular environment is highly crowded with biomacromolecules such as proteins and nucleic acids. Under such conditions, the structural and biophysical features of nucleic acids have been thought to be different from those in vitro. To obtain high-resolution...
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10-29-2020 10:21 AM
[NMR paper] In-Cell NMR in Human Cells: Direct Protein Expression Allows Structural Studies of Protein Folding and Maturation.
In-Cell NMR in Human Cells: Direct Protein Expression Allows Structural Studies of Protein Folding and Maturation.
In-Cell NMR in Human Cells: Direct Protein Expression Allows Structural Studies of Protein Folding and Maturation.
Acc Chem Res. 2018 Jun 05;:
Authors: Luchinat E, Banci L
Abstract
Cellular structural biology methods are needed to characterize biological processes at atomic resolution in the physiological environment of the cell. Toward this goal, solution in-cell NMR is a powerful approach because it provides...
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06-06-2018 01:42 PM
[NMR paper] Characterization of proteins by in-cell NMR spectroscopy in cultured mammalian cells.
Characterization of proteins by in-cell NMR spectroscopy in cultured mammalian cells.
Characterization of proteins by in-cell NMR spectroscopy in cultured mammalian cells.
Nat Protoc. 2016 Jun;11(6):1101-1111
Authors: Barbieri L, Luchinat E, Banci L
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05-20-2016 03:04 PM
[NMR paper] Sequential protein expression and selective labeling for in-cell NMR in human cells.
Sequential protein expression and selective labeling for in-cell NMR in human cells.
Related Articles Sequential protein expression and selective labeling for in-cell NMR in human cells.
Biochim Biophys Acta. 2015 Dec 23;
Authors: Luchinat E, Secci E, Cencetti F, Bruni P
Abstract
BACKGROUND: In-cell NMR is a powerful technique to investigate proteins in living human cells at atomic resolution. Ideally, when studying functional processes involving protein-protein interactions by NMR, only one partner should be isotopically...
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01-04-2016 07:49 PM
Sequential protein expression and selective labeling for in-cell NMR in human cells
Sequential protein expression and selective labeling for in-cell NMR in human cells
Publication date: Available online 23 December 2015
Source:Biochimica et Biophysica Acta (BBA) - General Subjects</br>
Author(s): Enrico Luchinat, Erica Secci, Francesca Cencetti, Paola Bruni</br>
Background In-cell NMR is a powerful technique to investigate proteins in living human cells at atomic resolution. Ideally, when studying functional processes involving protein-protein interactions by NMR, only one partner should be isotopically labeled. Here we show that...
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12-28-2015 12:26 AM
High-yield Escherichia coli-based cell-free expression of human proteins
High-yield Escherichia coli-based cell-free expression of human proteins
Abstract Production of sufficient amounts of human proteins is a frequent bottleneck in structural biology. Here we describe an Escherichia coli-based cell-free system which yields mg-quantities of human proteins in N-terminal fusion constructs with the GB1 domain, which show significantly increased translation efficiency. A newly generated E. coli BL21 (DE3) RIPL-Star strain was used, which contains a variant RNase E with reduced activity and an excess of rare-codon tRNAs, and is devoid of lon and ompT protease...
Direct Expression of Fluorinated Proteins in Human Cells for 19F In-Cell NMR Spectroscopy
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