Related ArticlesIn-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 disordered protein of alpha-Synuclein (?Syn) in mammalian cells. This chapter includes a simplified expression and purification protocol of recombinant ?Syn and its delivery into mammalian cells. The chapter also describes how to assess the cell leakage that might occur to the cells, the setup of the instrument, and how to perform basic analyses with the obtained NMR data.
[ASAP] How Do Intrinsically Disordered Viral Proteins Hijack the Cell?
How Do Intrinsically Disordered Viral Proteins Hijack the Cell?
https://pubs.acs.org/appl/literatum/publisher/achs/journals/content/bichaw/0/bichaw.ahead-of-print/acs.biochem.8b00622/20180627/images/medium/bi-2018-00622j_0003.gif
Biochemistry
DOI: 10.1021/acs.biochem.8b00622
http://feeds.feedburner.com/~ff/acs/bichaw?d=yIl2AUoC8zA
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06-29-2018 08:31 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
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05-20-2016 03:04 PM
[NMR paper] Algal autolysate medium to label proteins for NMR in mammalian cells.
Algal autolysate medium to label proteins for NMR in mammalian cells.
Related Articles Algal autolysate medium to label proteins for NMR in mammalian cells.
J Biomol NMR. 2016 Apr 22;
Authors: Fuccio C, Luchinat E, Barbieri L, Neri S, Fragai M
Abstract
In-cell NMR provides structural and functional information on proteins directly inside living cells. At present, the high costs of the labeled media for mammalian cells represent a limiting factor for the development of this methodology. Here we report a protocol to prepare a...
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04-24-2016 10:29 PM
Algal autolysate medium to label proteins for NMR in mammalian cells
Algal autolysate medium to label proteins for NMR in mammalian cells
Abstract
In-cell NMR provides structural and functional information on proteins directly inside living cells. At present, the high costs of the labeled media for mammalian cells represent a limiting factor for the development of this methodology. Here we report a protocol to prepare a homemade growth medium from Spirulina platensis autolysate, suitable to express uniformly labeled proteins inside mammalian cells at a reduced cost-per-sample. The human proteins SOD1 and Mia40 were...
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04-22-2016 08:45 PM
[NMR paper] In-cell (13)C NMR spectroscopy for the study of intrinsically disordered proteins.
In-cell (13)C NMR spectroscopy for the study of intrinsically disordered proteins.
In-cell (13)C NMR spectroscopy for the study of intrinsically disordered proteins.
Nat Protoc. 2014 Sep;9(9):2005-2016
Authors: Felli IC, Gonnelli L, Pierattelli R
Abstract
A large number of proteins carry out their function in highly flexible and disordered states, lacking a well-defined 3D structure. These proteins, referred to as intrinsically disordered proteins (IDPs), are now in the spotlight of modern structural biology. Nuclear magnetic...
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08-01-2014 06:21 PM
[NMR paper] Tyrosine Phosphorylation within the Intrinsically Disordered Cytosolic Domains of the B-Cell Receptor: An NMR-Based Structural Analysis.
Tyrosine Phosphorylation within the Intrinsically Disordered Cytosolic Domains of the B-Cell Receptor: An NMR-Based Structural Analysis.
Related Articles Tyrosine Phosphorylation within the Intrinsically Disordered Cytosolic Domains of the B-Cell Receptor: An NMR-Based Structural Analysis.
PLoS One. 2014;9(4):e96199
Authors: Rosenlöw J, Isaksson L, Mayzel M, Lengqvist J, Orekhov VY
Abstract
Intrinsically disordered proteins are found extensively in cell signaling pathways where they often are targets of posttranslational...
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04-29-2014 12:04 PM
[NMR paper] In-Cell NMR Characterization of the Secondary Structure Populations of a Disordered Conformation of ?-Synuclein within E. coli Cells.
In-Cell NMR Characterization of the Secondary Structure Populations of a Disordered Conformation of ?-Synuclein within E. coli Cells.
In-Cell NMR Characterization of the Secondary Structure Populations of a Disordered Conformation of ?-Synuclein within E. coli Cells.
PLoS One. 2013;8(8):e72286
Authors: Waudby CA, Camilloni C, Fitzpatrick AW, Cabrita LD, Dobson CM, Vendruscolo M, Christodoulou J
Abstract
?-Synuclein is a small protein strongly implicated in the pathogenesis of Parkinson's disease and related neurodegenerative disorders. We...
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08-31-2013 06:56 PM
[NMR paper] Highly efficient NMR assignment of intrinsically disordered proteins: application to B- and T cell receptor domains.
Highly efficient NMR assignment of intrinsically disordered proteins: application to B- and T cell receptor domains.
Related Articles Highly efficient NMR assignment of intrinsically disordered proteins: application to B- and T cell receptor domains.
PLoS One. 2013;8(5):e62947
Authors: Isaksson L, Mayzel M, Saline M, Pedersen A, Rosenlöw J, Brutscher B, Karlsson BG, Orekhov VY
Abstract
We present an integrated approach for efficient characterization of intrinsically disordered proteins. Batch cell-free expression, fast data acquisition,...
In-Cell NMR of Intrinsically Disordered Proteins in Mammalian Cells.
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