Issue 4, 2015

Probing coiled-coil assembly by paramagnetic NMR spectroscopy

Abstract

Here a new method to determine the oligomeric state and orientation of coiled-coil peptide motifs is described. Peptides K and E, which are designed to form a parallel heterodimeric complex in aqueous solution, were labeled with the aromatic amino acids tryptophan and tyrosine on the C-terminus respectively as ‘fingerprint’ residues. One of the peptides was also labeled with the paramagnetic probe MTSL. One dimensional proton NMR spectroscopy was used to study the peptide quaternary structure by monitoring the signal suppression of the aromatic labels due to proximity of the nitroxyl radical. 1D-NMR confirmed that the peptides K and E form a heterodimeric coiled coil with a parallel orientation. In addition, fluorescence emission quenching of the aromatic labels due to electron exchange with a nitroxyl radical confirmed the parallel coiled coil orientation. Thus, paramagnetic nitroxide and aromatic fluorophore labeling of peptides yields valuable information regarding the quaternary structure from 1D-NMR and steady-state fluorescence measurements. This convenient method is useful not only to investigate coiled coil assembly, but can also be applied to any defined supramolecular assembly.

Graphical abstract: Probing coiled-coil assembly by paramagnetic NMR spectroscopy

Supplementary files

Article information

Article type
Paper
Submitted
05 אוק 2014
Accepted
17 נוב 2014
First published
18 נוב 2014
This article is Open Access
Creative Commons BY-NC license

Org. Biomol. Chem., 2015,13, 1159-1168

Author version available

Probing coiled-coil assembly by paramagnetic NMR spectroscopy

T. Zheng, A. Boyle, H. Robson Marsden, D. Valdink, G. Martelli, J. Raap and A. Kros, Org. Biomol. Chem., 2015, 13, 1159 DOI: 10.1039/C4OB02125H

This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence. You can use material from this article in other publications, without requesting further permission from the RSC, provided that the correct acknowledgement is given and it is not used for commercial purposes.

To request permission to reproduce material from this article in a commercial publication, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party commercial publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements