NMR characterization of the C-mannose conformation in a thrombospondin repeat using a selective labeling approach.
Angew Chem Int Ed Engl. 2020 Aug 03;:
Authors: Jonker HRA, Saxena K, Shcherbakova A, Tiemann B, Bakker H, Schwalbe H
Abstract
Despite the great interest in glycoproteins, structural information reporting on conformation and dynamics of the sugar moieties are limited. We present a new biochemical method to express proteins with glycans that are selectively labeled with NMR active nuclei. We report on the incorporation of 13C-labeled mannose in the C-mannosylated UNC-5 thrombospondin repeat. The conformational landscape of the C-mannose sugar puckers attached to tryptophan residues of UNC-5 is characterized by interconversion between the canonical 1C4 state and the B03 / 1S3 state. This flexibility may be essential for protein folding and stabilization. We foresee that this versatile tool to produce proteins with selective labeled C-mannose can be applied and adjusted to other systems and modifications and potentially paves a way to advance glycoprotein research by unravelling the dynamical and conformational properties of glycan structures and their interactions.
PMID: 32745319 [PubMed - as supplied by publisher]
Identification of conformation-selective nanobodies against the membrane protein insertase BamA by an integrated structural biology approach
Identification of conformation-selective nanobodies against the membrane protein insertase BamA by an integrated structural biology approach
Abstract
The insertase BamA is an essential protein of the bacterial outer membrane. Its 16-stranded transmembrane β-barrel contains a lateral gate as a key functional element. This gate is formed by the C-terminal half of the last β-strand. The BamA barrel was previously found to sample different conformations in aqueous solution, as well as different gate-open, gate-closed, and collapsed conformations in...
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05-09-2019 05:55 AM
[NMR paper] A Combined NMR and Computational Approach to Investigate Peptide Binding to a Designed Armadillo Repeat Protein.
A Combined NMR and Computational Approach to Investigate Peptide Binding to a Designed Armadillo Repeat Protein.
A Combined NMR and Computational Approach to Investigate Peptide Binding to a Designed Armadillo Repeat Protein.
J Mol Biol. 2015 Mar 24;
Authors: Ewald C, Christen MT, Watson RP, Mihajlovic M, Zhou T, Honegger A, Plückthun A, Caflisch A, Zerbe O
Abstract
The specific recognition of peptide sequences by proteins plays an important role both in biology and in diagnostic applications. Here we characterize the...
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03-31-2015 07:17 PM
A Combined NMR and Computational Approach to Investigate Peptide Binding to a Designed Armadillo Repeat Protein
A Combined NMR and Computational Approach to Investigate Peptide Binding to a Designed Armadillo Repeat Protein
Publication date: Available online 24 March 2015
Source:Journal of Molecular Biology</br>
Author(s): Christina Ewald , Martin T. Christen , Randall P. Watson , Maja Mihajlovic , Ting Zhou , Annemarie Honegger , Andreas Plückthun , Amedeo Caflisch , Oliver Zerbe</br>
The specific recognition of peptide sequences by proteins plays an important role both in biology and in diagnostic applications. Here we characterize the relatively weak binding of the...
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03-25-2015 10:15 AM
Frequency-selective heteronuclear dephasing and selective carbonyl labeling to deconvolute crowded spectra of membrane proteins by magic angle spinning NMR.
Frequency-selective heteronuclear dephasing and selective carbonyl labeling to deconvolute crowded spectra of membrane proteins by magic angle spinning NMR.
Frequency-selective heteronuclear dephasing and selective carbonyl labeling to deconvolute crowded spectra of membrane proteins by magic angle spinning NMR.
J Magn Reson. 2011 Mar 17;
Authors: Traaseth NJ, Veglia G
We present a new method that combines carbonyl-selective labeling with frequency-selective heteronuclear recoupling to resolve the spectral overlap of magic angle spinning (MAS) NMR...
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04-13-2011 11:57 PM
Frequency-Selective Heteronuclear Dephasing and Selective Carbonyl Labeling to Deconvolute Crowded Spectra of Membrane Proteins By Magic Angle Spinning NMR
Frequency-Selective Heteronuclear Dephasing and Selective Carbonyl Labeling to Deconvolute Crowded Spectra of Membrane Proteins By Magic Angle Spinning NMR
Publication year: 2011
Source: Journal of Magnetic Resonance, In Press, Accepted Manuscript, Available online 17 March 2011</br>
Nathaniel J., Traaseth , Gianluigi, Veglia</br>
We present a new method that combines carbonyl-selective labeling with frequency-selective heteronuclear recoupling to resolve the spectral overlap of magic angle spinning (MAS) NMR spectra of membrane proteins in fluid lipid membranes with broad lines and...
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03-18-2011 06:43 AM
[NMR paper] Site-selective labeling strategies for screening by NMR.
Site-selective labeling strategies for screening by NMR.
Related Articles Site-selective labeling strategies for screening by NMR.
Comb Chem High Throughput Screen. 2002 Dec;5(8):623-30
Authors: Weigelt J, Wikström M, Schultz J, van Dongen MJ
NMR based screening has become an important tool in the pharmaceutical industry. Methods that provide information on the location of small molecule binding sites on the surface of a drug target (e. g. SAR-by-NMR and related techniques) are of particular interest. In order to extend the applicability of...
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11-24-2010 08:58 PM
[NMR paper] A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein r
A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein revealed by NMR spectroscopy.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein revealed by NMR spectroscopy.
FEBS Lett. 1997 Jun 9;409(2):227-31
Authors: Tsuda S, Ito A, Matsushima N
The 1H-NMR spectrum of a synthetic 24-residue peptide (A1-G-V-D-S-S-L-I-A-G-Y-G-S-T-Q-T-S-G-S-D-S-A-L-T24; INP24),...
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[NMR paper] A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein r
A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein revealed by NMR spectroscopy.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein revealed by NMR spectroscopy.
FEBS Lett. 1997 Jun 9;409(2):227-31
Authors: Tsuda S, Ito A, Matsushima N
The 1H-NMR spectrum of a synthetic 24-residue peptide (A1-G-V-D-S-S-L-I-A-G-Y-G-S-T-Q-T-S-G-S-D-S-A-L-T24; INP24),...
NMR characterization of the C-mannose conformation in a thrombospondin repeat using a selective labeling approach.
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