Related ArticlesRecording In-Cell NMR-Spectra in Living Mammalian Cells.
Methods Mol Biol. 2020;2141:857-871
Authors: Mate?ko-Burmann I, Burmann BM
Abstract
At the foundation of many cellular processes as well as a large number of diseases is the (mis)folding of important intrinsically disordered proteins (IDPs). Despite tremendous scientific efforts, the factors driving their structural changes within the cellular context remain poorly understood. In-cell NMR spectroscopy enables investigation of IDPs directly in the living eukaryotic cell enabling investigation of its intermolecular interactions and ensuing modifications at an unprecedented atomic resolution. In the following protocol, we describe how to prepare in-cell NMR samples of IDPs within eukaryotic cells and how to measure these in-cell NMR samples of an IDP in its natural environment, the living mammalian cell. Furthermore, we outline a procedure to assess the intracellular recombinant protein concentration of the studied IDP based on in-cell NMR methods. We use ?-synuclein as a model protein, but the presented approach is highly modular and therefore should be easily adapted and altered to the desired needs for the studies of different IDPs.
[NMR paper] In-Cell NMR of Intrinsically Disordered Proteins in Mammalian Cells.
In-Cell NMR of Intrinsically Disordered Proteins in Mammalian Cells.
Related Articles In-Cell NMR of Intrinsically Disordered Proteins in Mammalian Cells.
Methods Mol Biol. 2020;2141:873-893
Authors: Gerez JA, Prymaczok NC, Riek R
Abstract
In-cell NMR enables structural insights at atomic resolution of proteins in their natural environment. To date, very few methods have been developed to study proteins by in-cell NMR in mammalian systems. Here we describe a detailed protocol to conduct in-cell NMR on the intrinsically...
nmrlearner
Journal club
0
07-23-2020 11:23 PM
[NMR paper] Recent progress of in-cell NMR of nucleic acids in living human cells.
Recent progress of in-cell NMR of nucleic acids in living human cells.
Related Articles Recent progress of in-cell NMR of nucleic acids in living human cells.
Biophys Rev. 2020 Mar 06;:
Authors: Yamaoki Y, Nagata T, Sakamoto T, Katahira M
Abstract
The inside of living cells is highly crowded with biological macromolecules. It has long been considered that the properties of nucleic acids and proteins, such as their structures, dynamics, interactions, and enzymatic activities, in intracellular environments are different from those...
nmrlearner
Journal club
0
03-09-2020 01:12 PM
[NMR paper] The first successful observation of in-cell NMR signals of DNA and RNA in living human cells.
The first successful observation of in-cell NMR signals of DNA and RNA in living human cells.
Related Articles The first successful observation of in-cell NMR signals of DNA and RNA in living human cells.
Phys Chem Chem Phys. 2017 Oct 12;:
Authors: Yamaoki Y, Kiyoishi A, Miyake M, Kano F, Murata M, Nagata T, Katahira M
Abstract
In order to understand intracellular biological events, information on the structure, dynamics and interaction of proteins and nucleic acids in living cells is of crucial importance. In-cell NMR is a...
nmrlearner
Journal club
0
10-14-2017 02:04 AM
[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
Abstract
nmrlearner
Journal club
0
05-20-2016 03:04 PM
[NMR paper] Fluorine NMR-based assay in living mammalian cells.
Fluorine NMR-based assay in living mammalian cells.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Fluorine NMR-based assay in living mammalian cells.
Anal Biochem. 2015 Dec 11;
Authors: Veronesi M, Giacomina F, Romeo E, Castellani B, Ottonello G, Lambruschini C, Garau G, Scarpelli R, Bandiera T, Piomelli D, Dalvit C
Abstract
Nuclear Magnetic Resonance (NMR)-based screening has been recognized as a powerful approach for the identification...
nmrlearner
Journal club
0
12-28-2015 12:26 AM
[NMR paper] Protein dynamics in living cells studied by in-cell NMR spectroscopy.
Protein dynamics in living cells studied by in-cell NMR spectroscopy.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Protein dynamics in living cells studied by in-cell NMR spectroscopy.
FEBS Lett. 2013 Jan 11;
Authors: Li C, Liu M
Abstract
Most proteins function in cells where protein concentrations can reach 400g/l. However, most quantitative studies of protein properties are performed in idealized, dilute conditions. Recently developed in-cell NMR techniques...
nmrlearner
Journal club
0
02-03-2013 10:22 AM
Protein dynamics in living cells studied by in-cell NMR spectroscopy
Protein dynamics in living cells studied by in-cell NMR spectroscopy
Available online 11 January 2013
Publication year: 2013
Source:FEBS Letters</br>
</br>
Most proteins function in cells where protein concentrations can reach 400g/l. However, most quantitative studies of protein properties are performed in idealized, dilute conditions. Recently developed in-cell NMR techniques can provide protein structure and other biophysical properties inside living cells at atomic resolution. Here we review how protein dynamics, including global and internal motions have been...
nmrlearner
Journal club
0
02-03-2013 10:13 AM
On-Cell MAS NMR: PhysiologicalClues from Living Cells
On-Cell MAS NMR: PhysiologicalClues from Living Cells
Giorgia Zandomeneghi, Karin Ilg, Markus Aebi and Beat H. Meier
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja307467p/aop/images/medium/ja-2012-07467p_0007.gif
Journal of the American Chemical Society
DOI: 10.1021/ja307467p
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/PBl2HVolQkY
Recording In-Cell NMR-Spectra in Living Mammalian Cells.
Linear Mode
