Skip Navigation


PEDS Advance Access originally published online on March 14, 2006
Protein Engineering Design and Selection 2006 19(5):211-217; doi:10.1093/protein/gzl003
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow All Versions of this Article:
19/5/211    most recent
gzl003v1
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 ISI Web of Science
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 arrow Search for citing articles in:
ISI Web of Science (3)
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Park, S.
Right arrow Articles by Boder, E. T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Park, S.
Right arrow Articles by Boder, E. T.
Related Collections
Right arrow 2006
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

Limitations of yeast surface display in engineering proteins of high thermostability

Sheldon Park1, Yao Xu1, Xiaoran Fu Stowell1,4, Feng Gai1,3, Jeffery G. Saven1,3 and Eric T. Boder2,3,5

1Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104, 2Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, PA 19104 and 3Laboratory for Research on the Structure of Matter, University of Pennsylvania, Philadelphia, PA 19104, USA 4Present address: Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA

5 To whom correspondence should be addressed. E-mail: boder{at}seas.upenn.edu

Engineering proteins that can fold to unique structures remains a challenge. Protein stability has previously been engineered via the observed correlation between thermal stability and eukaryotic secretion level. To explore the limits of an expression-based approach, variants of the highly thermostable three-helix bundle protein {alpha}3D were studied using yeast surface display. A library of {alpha}3D mutants was created to explore the possible correlation of protein stability and fold with expression level. Five efficiently expressed mutants were then purified and further studied biochemically. Despite their differences in stability, most mutants expressed at levels comparable with that of wild-type {alpha}3D. Two other related sequences ({alpha}3A and {alpha}3B) that form collapsed, stable molten globules but lack a uniquely folded structure were similarly expressed at high levels by yeast display. Together these observations suggest that the quality control system in yeast is unable to discriminate between well-folded proteins of high stability and molten globules. The present study, therefore, suggests that an optimization of the surface display efficiency on yeast may yield proteins that are thermally and chemically stable yet are poorly folded.

Keywords: {alpha}3D/helical protein/protein library/yeast surface display

Received July 15, 2005; revised December 2, 2005; accepted January 30, 2006.

Edited by James Marks


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
 Protein Eng Des SelHome page
C. Kiss, J. Temirov, L. Chasteen, G. S. Waldo, and A. R.M. Bradbury
Directed evolution of an extremely stable fluorescent protein
Protein Eng. Des. Sel., May 1, 2009; 22(5): 313 - 323.
[Abstract] [Full Text] [PDF]

Home page
 BioinformaticsHome page
M. Fromer and C. Yanover
A computational framework to empower probabilistic protein design
Bioinformatics, July 1, 2008; 24(13): i214 - i222.
[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.