Skip Navigation



PEDS Advance Access published online on May 23, 2006
Protein Engineering Design and Selection, doi:10.1093/protein/gzl018
This Article
Right arrow FREE Full Text (PDF) Freely available
Right arrow All Versions of this Article:
19/8/385    most recent
gzl018v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Schlapschy, M.
Right arrow Articles by Skerra, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Schlapschy, M.
Right arrow Articles by Skerra, A.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2006. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org
Received March 28, 2006
Accepted April 14, 2006

Short Communication

A system for concomitant overexpression of four periplasmic folding catalysts to improve secretory protein production in Escherichia coli

Martin Schlapschy 1, Sebastian Grimm 1, and Arne Skerra 1 *

1 Lehrstuhl für Biologische Chemie, Technische Universität München, 85350 Freising-Weihenstephan, Germany

* To whom correspondence should be addressed.
Arne Skerra, E-mail: skerra{at}wzw.tum.de


  Abstract

Although Escherichia coli is in wide use for preparative protein expression, problems with the folding of the recombinant gene product and protein aggregation are frequently encountered. Apart from cytoplasmic expression, this is also true for secretion into the bacterial periplasm, the method of choice for the production of proteins that carry structural disulfide bonds. Here we report the construction of the helper plasmid pTUM4, which effects overexpression of four established periplasmic chaperones and folding catalysts: the thiol-disulfide oxidoreductases DsbA and DsbC that catalyze the formation and isomerization of disulfide bridges and the peptidyl-prolyl cis/trans-isomerases with chaperone activity, FkpA and SurA. pTUM4 carries a p15a origin of replication and a chloramphenicol resistance gene and, thus, it is compatible with many conventional expression vectors that use the ColEI origin and an ampicillin resistance. Its positive effects on the yield of soluble recombinant protein and the homogeneity of disulfide pattern are illustrated here using the human plasma retinol-binding protein as well as the extracellular carbohydrate recognition domain of the dendritic cell membrane receptor DC-SIGN. Hence, pTUM4 represents a novel helper vector which complements existing cytosolic chaperone coexpression plasmids and should be useful for the functional secretion of various recombinant proteins with hampered folding efficiency.

Keywords: chaperone; disulfide isomerase; DsbA; DsbC; FkpA; folding catalyst; peptidyl-prolyl cis; trans-isomerase; SurA.
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Proc. Natl. Acad. Sci. USAHome page
N. S. Outchkourov, W. Roeffen, A. Kaan, J. Jansen, A. Luty, D. Schuiffel, G. J. van Gemert, M. van de Vegte-Bolmer, R. W. Sauerwein, and H. G. Stunnenberg
Correctly folded Pfs48/45 protein of Plasmodium falciparum elicits malaria transmission-blocking immunity in mice
PNAS, March 18, 2008; 105(11): 4301 - 4305.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.