PuSH - Publication Server of Helmholtz Zentrum München: Effects of fluorophore attachment on protein conformation and dynamics studied by spFRET and NMR spectroscopy.

Navigation

Home

Deutsch

Research

Advanced Search

Browse by ...

... Journal

... Publication Type

... Research Data

... Publication Year

Publication overview

Support & Contact

Contact persons

Help

Data protection

Sanchez-Rico, C. ; VoithvonVoithenberg, L.* ; Warner, L. ; Lamb, D.C.* ; Sattler, M.

Effects of fluorophore attachment on protein conformation and dynamics studied by spFRET and NMR spectroscopy.

Chem. Eur. J. 23, 14267-14277 (2017)

DOI

PMC

Open Access Green as soon as Postprint is submitted to ZB.

Abstract
Metrics
Extra information

Fluorescence-based techniques are widely used to study biomolecular conformations, intra- and intermolecular interactions, and conformational dynamics of macromolecules. Especially for fluorescence-based single-molecule experiments, the choice of the fluorophore and labeling position are highly important. In this work, we studied the biophysical and structural effects that are associated with the conjugation of fluorophores to cysteines in the splicing factor U2AF65 by using single pair Förster resonance energy transfer (FRET) and nuclear magnetic resonance (NMR) spectroscopy. It is shown that certain acceptor fluorophores are advantageous depending on the experiments performed. The effects of dye attachment on the protein conformation were characterized using heteronuclear NMR experiments. The presence of hydrophobic and aromatic moieties in the fluorophores can significantly affect the conformation of the conjugated protein, presumably by transient interactions with the protein surface. Guidelines are provided for carefully choosing fluorophores, considering their photophysical properties and chemical features for the design of FRET experiments, and for minimizing artifacts.

Altmetric

Additional Metrics?

[➜Log in]

Edit extra informations Login

Publication type Article: Journal article

Document type Scientific Article

Corresponding Author

Keywords Fluorophores ; Fret ; Multidomain Proteins ; Nmr Spectroscopy ; Protein-rna Interactions; Single-molecule Fret; Dyes Intramolecular Stabilization; Spin Labels; Organic Fluorophores; Fluorescent Proteins; Energy-transfer; Cell; Probes; Dna; Microscopy

ISSN (print) / ISBN 0947-6539

e-ISSN 1521-3765

Journal Chemistry - A European Journal

Quellenangaben Volume: 23, Issue: 57, Pages: 14267-14277

Publisher Wiley

Publishing Place Weinheim

Non-patent literature Publications

Reviewing status Peer reviewed

Institute(s) Institute of Structural Biology (STB)