Reconstitution of Isotopically Labeled Ribosomal Protein L29 in the 50S Large Ribosomal Subunit for Solution-State and Solid-State NMR.
Methods Mol Biol. 2018;1764:87-100
Authors: Barbet-Massin E, van der Sluis E, Musial J, Beckmann R, Reif B
Abstract
Solid-state nuclear magnetic resonance (NMR) has recently emerged as a method of choice to study structural and dynamic properties of large biomolecular complexes at atomic resolution. Indeed, recent technological and methodological developments have enabled the study of ever more complex systems in the solid-state. However, to explore multicomponent protein complexes by NMR, specific labeling schemes need to be developed that are dependent on the biological question to be answered. We show here how to reconstitute an isotopically labeled protein within the unlabeled 50S or 70S ribosomal subunit. In particular, we focus on the 63-residue ribosomal protein L29 (~7*kDa), which is located at the exit of the tunnel of the large 50S ribosomal subunit (~1.5*MDa). The aim of this work is the preparation of a suitable sample to investigate allosteric conformational changes in a ribosomal protein that are induced by the nascent polypeptide chain and that trigger the interaction with different chaperones (e.g., trigger factor or SRP).
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Angew Chem Int Ed Engl. 2015 Feb 5;
Authors: Barbet-Massin E, Huang CT, Daebel V, Hsu ST, Reif B
Abstract
Co-translational protein folding is not yet well understood despite the availability of high-resolution ribosome crystal structures. We present first solid-state NMR data on non-mobile regions of a...
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[NMR paper] Strategies for solid-state NMR investigations of supramolecular assemblies with large subunit sizes.
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J Magn Reson. 2014 Nov 15;
Authors: Fricke P, Chevelkov V, Shi C, Lange A
Abstract
Solid-state NMR is a versatile tool to study structure and dynamics of insoluble and non-crystalline biopolymers. Supramolecular protein assemblies are formed by self-association of multiple copies of single small-sized...
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[NMR paper] Strategies for solid-state NMR investigations of supramolecular assemblies with large subunit sizes
Strategies for solid-state NMR investigations of supramolecular assemblies with large subunit sizes
Publication date: Available online 15 November 2014
Source:Journal of Magnetic Resonance</br>
Author(s): Pascal Fricke , Veniamin Chevelkov , Chaowei Shi , Adam Lange</br>
Solid-state NMR is a versatile tool to study structure and dynamics of insoluble and non-crystalline biopolymers. Supramolecular protein assemblies are formed by self-association of multiple copies of single small-sized proteins. Because of their high degree of local order, solid-state NMR...
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11-16-2014 02:13 AM
[NMR paper] Preparation of uniformly isotope labeled KcsA for solid state NMR: Expression, purification, reconstitution into liposomes and functional assay.
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Protein Expr Purif. 2013 Aug 1;
Authors: Bhate MP, Wylie BJ, Thompson A, Tian L, Nimigean C, McDermott AE
Abstract
We report the expression, purification, liposome reconstitution and functional validation of uniformly (13)C and (15)N isotope labeled...
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[NMR paper] The NMR solution structure of the 30S ribosomal protein S27e encoded in gene RS27_ARC
The NMR solution structure of the 30S ribosomal protein S27e encoded in gene RS27_ARCFU of Archaeoglobus fulgidis reveals a novel protein fold.
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Protein Sci. 2004 May;13(5):1407-16
Authors: Herve du Penhoat C, Atreya HS, Shen Y, Liu G, Acton TB, Xiao R, Li Z, Murray D, Montelione GT, Szyperski T
The Archaeoglobus fulgidis gene RS27_ARCFU encodes the 30S ribosomal protein S27e. Here, we...
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11-24-2010 09:51 PM
[NMR paper] Solution NMR structure of the 30S ribosomal protein S28E from Pyrococcus horikoshii.
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We report NMR assignments and solution structure of the 71-residue 30S ribosomal protein S28E from the archaean Pyrococcus horikoshii, target JR19 of the Northeast Structural...
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Int J Pept Protein Res. 1996 Apr;47(4):282-8
Authors: Katahira R, Flotow H, Thomas G, Nosaka AY
An increase in the rate of protein synthesis is found to be accompanied by phosphorylation of the 40S ribosomal protein S6. Treatment of S6 by cyanogen bromide produced three fragments, and one of the fragments of S6, which is a C-terminal...
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[NMR paper] Solution structure of prokaryotic ribosomal protein S17 by high-resolution NMR spectr
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http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--pubs.acs.org-images-acspubs.jpg Related Articles Solution structure of prokaryotic ribosomal protein S17 by high-resolution NMR spectroscopy.
Biochemistry. 1996 Mar 5;35(9):2845-53
Authors: Jaishree TN, Ramakrishnan V, White SW
The solution of a primary 16S rRNA-binding ribosomal protein, S17, was investigated by two- and three-dimensional homonuclear and heteronuclear magnetic resonance...
Reconstitution of Isotopically Labeled Ribosomal Protein L29 in the 50S Large Ribosomal Subunit for Solution-State and Solid-State NMR.
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