Advanced approaches for the characterization of a de novo designed antiparallel coiled coil peptide

Kevin Pagel; Karsten Seeger; Bettina Seiwert; Alessandra Villa; Alan E. Mark; Stefan Berger; Beate Koksch

doi:10.1039/B418167K

Advanced approaches for the characterization of a de novo designed antiparallel coiled coil peptide

Kevin Pagel,^a Karsten Seeger,^b Bettina Seiwert,^b Alessandra Villa,†^c Alan E. Mark,^c Stefan Berger^b and Beate Koksch*^a

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* Corresponding authors

^a Free University Berlin, Department of Chemistry—Organic Chemistry, Takustrasse 3, Berlin, Germany
E-mail: koksch@chemie.fu-berlin.de
Fax: +49-30-838-55644
Tel: +49-30-838-55386

^b University of Leipzig, Department of Analytical Chemistry, Linnestrasse 3, Leipzig, Germany
E-mail: stberger@rz.uni-leipzig.de
Fax: +49-341-9711833
Tel: +49-341-9736101

^c Department of Biophysical Chemistry, University of Groningen, Nijenborgh 4, Groningen, The Netherlands
E-mail: a.e.mark@rug.nl
Fax: + 31-50-363-4800
Tel: +31-50-363-4457

Abstract

We report here an advanced approach for the characterization of the folding pattern of a de novo designed antiparallel coiled coil peptide by high-resolution methods. Incorporation of two fluorescence labels at the C- and N-terminus of the peptide chain as well as modification of two hydrophobic core positions by Phe/[¹⁵N,¹³C]Leu enable the study of the folding characteristics and of distinct amino acid side chain interactions by fluorescence resonance energy transfer (FRET) and NMR spectroscopy. Results of both experiments reveal the antiparallel alignment of the helices and thus prove the design concept. This finding is also supported by molecular dynamics simulations. Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR-MS) in combination with NMR experiments was used for verification of the oligomerization equilibria of the coiled coil peptide.

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https://doi.org/10.1039/B418167K

Article type

Paper

Submitted

02 Dec 2004

Accepted

07 Feb 2005

First published

24 Feb 2005

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Org. Biomol. Chem., 2005,3, 1189-1194

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Advanced approaches for the characterization of a de novo designed antiparallel coiled coil peptide

K. Pagel, K. Seeger, B. Seiwert, A. Villa, A. E. Mark, S. Berger and B. Koksch, Org. Biomol. Chem., 2005, 3, 1189 DOI: 10.1039/B418167K

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Organic & Biomolecular Chemistry

Advanced approaches for the characterization of a de novo designed antiparallel coiled coil peptide

Abstract

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Advanced approaches for the characterization of a de novo designed antiparallel coiled coil peptide

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