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

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

249,00 € / $374.00 / £187.00*

See all formats and pricing



Select Volume and Issue
Loading journal volume and issue information...

Efficient production of native actin upon translation in a bacterial lysate supplemented with the eukaryotic chaperonin TRiC

Markus J. Stemp1 / Suranjana Guha2 / F. Ulrich Hartl3 / José M. Barral4





Corresponding author

Citation Information: Biological Chemistry. Volume 386, Issue 8, Pages 753–757, ISSN (Online) 1437-4315, ISSN (Print) 1431-6730, DOI: 10.1515/BC.2005.088, September 2005

Publication History

March 16, 2005
April 7, 2005
Published Online:


Recombinant expression of actin in bacteria results in non-native species that aggregate into inclusion bodies. Actin is a folding substrate of TRiC, the chaperonin of the eukaryotic cytosol. By employing bacterial in vitro translation lysates supplemented with purified chaperones, we have found that TRiC is the only eukaryotic chaperone necessary for correct folding of newly translated actin. The actin thus produced binds deoxyribonuclease I and polymerizes into filaments, hallmarks of its native state. In contrast to its rapid folding in the eukaryotic cytosol, actin translated in TRiC-supplemented bacterial lysate folds with slower kinetics, resembling the kinetics upon refolding from denaturant. Lysate supplementation with the bacterial chaperonin GroEL/ES or the DnaK/DnaJ/GrpE chaperones leads to prevention of actin aggregation, yet fails to support its correct folding. This combination of in vitro bacterial translation and TRiC-assisted folding allows a detailed analysis of the mechanisms necessary for efficient actin folding in vivo. In addition, it provides a robust alternative for the production of substantial amounts of eukaryotic proteins that otherwise misfold or lead to cellular toxicity upon expression in heterologous hosts.

Keywords: molecular chaperones; protein folding; recombinant expression

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.

F. Russmann, M. J. Stemp, L. Monkemeyer, S. A. Etchells, A. Bracher, and F. U. Hartl
Proceedings of the National Academy of Sciences, 2012, Volume 109, Number 52, Page 21208
G. M Altschuler and K. R Willison
Journal of The Royal Society Interface, 2008, Volume 5, Number 29, Page 1391
Günter Pappenberger, Elizabeth A. McCormack, and Keith R. Willison
Journal of Molecular Biology, 2006, Volume 360, Number 2, Page 484
Julie Grantham, Ingrid Lassing, and Roger Karlsson
Protoplasma, 2012, Volume 249, Number 4, Page 1001
Annette Haacke, Gabriele Fendrich, Paul Ramage, and Martin Geiser
Protein Expression and Purification, 2009, Volume 64, Number 2, Page 185
Christian Behrends, Carola A. Langer, Raina Boteva, Ulrike M. Böttcher, Markus J. Stemp, Gregor Schaffar, Bharathi Vasudeva Rao, Armin Giese, Hans Kretzschmar, Katja Siegers, and F. Ulrich Hartl
Molecular Cell, 2006, Volume 23, Number 6, Page 887
Efraín Siller, Diane C. DeZwaan, John F. Anderson, Brian C. Freeman, and José M. Barral
Journal of Molecular Biology, 2010, Volume 396, Number 5, Page 1310
Derrick Kok Sing Tay, Gobinath Rajagopalan, Xiang Li, Yu Chen, Linda H.L. Lua, and Susanna Su Jan Leong
Biotechnology and Bioengineering, 2011, Volume 108, Number 3, Page 572
Karen I. Brackley and Julie Grantham
Cell Stress and Chaperones, 2009, Volume 14, Number 1, Page 23

Comments (0)

Please log in or register to comment.