Jump to ContentJump to Main Navigation
Show Summary Details

Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board Member: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Sies, Helmut / Turk, Boris / Wittinghofer, Alfred

12 Issues per year


IMPACT FACTOR 2015: 2.710
Rank 142 out of 289 in category Biochemistry & Molecular Biology in the 2015 Thomson Reuters Journal Citation Report/Science Edition

SCImago Journal Rank (SJR) 2015: 1.607
Source Normalized Impact per Paper (SNIP) 2015: 0.751
Impact per Publication (IPP) 2015: 2.609

Online
ISSN
1437-4315
See all formats and pricing

 


 
 

Select Volume and Issue

Issues

Applicability of superfolder YFP bimolecular fluorescence complementation in vitro

Corinna Ottmann
  • 1Department of Structural Biology, Max Planck Institute of Molecular Physiology, Otto-Hahn-Str. 11, D-44227 Dortmund, Germany
/ Michael Weyand
  • 2Chemical Genomics Centre of the Max Planck Society, Otto-Hahn-Str. 15, D-44227 Dortmund, Germany
/ Alexander Wolf
  • 3Chemical Genomics Centre of the Max Planck Society, Otto-Hahn-Str. 15, D-44227 Dortmund, Germany
/ Jürgen Kuhlmann
  • 4Department of Structural Biology, Max Planck Institute of Molecular Physiology, Otto-Hahn-Str. 11, D-44227 Dortmund, Germany
/ Christian Ottmann
  • 5Chemical Genomics Centre of the Max Planck Society, Otto-Hahn-Str. 15, D-44227 Dortmund, Germany
Published Online: 2008-11-13 | DOI: https://doi.org/10.1515/BC.2009.008

Abstract

Bimolecular fluorescence complementation (BiFC) using yellow fluorescent protein (YFP) is a widely employed method to study protein-protein interactions in cells. As yet, this technique has not been used in vitro. To evaluate a possible application of BiFC in vitro, we constructed a ‘superfolder split YFP’ system where 15 mutations enhance expression of the fusion proteins in Escherichia coli and enable a native purification due to improved solubility. Here, we present the crystal structure of ‘superfolder YFP’, providing the structural basis for the enhanced folding and stability characteristics. Complementation between the two non-fluorescent YFP fragments fused to HRas and Raf1RBD or to 14-3-3 and PMA2-CT52 resulted in the constitution of the functional fluorophore. The in vivo BiFC with these protein interaction pairs was demonstrated in eukaryotic cell lines as well. Here, we present for the first time BiFC in vitro studies with natively purified superfolder YFP fusion proteins and show the potential and drawbacks of this method for analyzing protein-protein interactions.

Keywords: 14-3-3; BiFC; fusicoccin; protein-protein interactions; Raf; Ras

Corresponding author


Received: 2008-09-04

Accepted: 2008-10-01

Published Online: 2008-11-13

Published in Print: 2009-01-01


Citation Information: Biological Chemistry. Volume 390, Issue 1, Pages 81–90, ISSN (Online) 1437-4315, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/BC.2009.008, November 2008

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

[1]
Xiaohong Liu, Jiasong Li, Cheng Hu, Qing Zhou, Wei Zhang, Meirong Hu, Juanzuo Zhou, and Jiangyun Wang
Angewandte Chemie, 2013, Volume 125, Number 18, Page 4905
[3]
Masami Isogai, Yoshihiro Kawamoto, Kazuto Inahata, Harumi Fukada, Kenji Sugimoto, and Toshiji Tada
Bioorganic & Medicinal Chemistry Letters, 2011, Volume 21, Number 10, Page 3021
[4]
Rachel H Rose, Stephen J Briddon, and Nicholas D Holliday
British Journal of Pharmacology, 2010, Volume 159, Number 4, Page 738
[5]
Shang-Te Danny Hsu, Georg Blaser, and Sophie E. Jackson
Chemical Society Reviews, 2009, Volume 38, Number 10, Page 2951

Comments (0)

Please log in or register to comment.