Skip Navigation



PEDS Advance Access published online on September 26, 2006
Protein Engineering Design and Selection, doi:10.1093/protein/gzl039
This Article
Right arrow FREE Full Text (PDF) Freely available
Right arrow All Versions of this Article:
19/11/517    most recent
gzl039v1
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 DiTursi, M. K.
Right arrow Articles by Dordick, J. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by DiTursi, M. K.
Right arrow Articles by Dordick, J. S.
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 April 28, 2006
Revised July 24, 2006
Accepted August 18, 2006

Article

Bioinformatics-driven, rational engineering of protein thermostability

Mary Kate DiTursi 1, Seok-Joon Kwon 1, Philippa J. Reeder 1, and Jonathan S. Dordick 1 *

1 Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180-3590, USA

* To whom correspondence should be addressed.
Jonathan S. Dordick, E-mail: dordick{at}rpi.edu


  Abstract

A longstanding goal in protein engineering is to identify specific sequence changes that endow proteins with desired functional properties. As opposed to traditional rational and random protein engineering techniques, we have employed a bioinformatic approach to identify specific sequence changes that influence key functional properties of a protein within a defined superfamily. Specifically, we have used the Bayesian sequence-based algorithms PROBE and Classifier to identify a strand-turn-strand motif that contributes to thermophilicity among members of the serine protease subtilase superfamily. By replacing a 16 amino acid sequence in the mesophilic subtilisin E (from Bacillus subtilis) with a bioinformatics-generated thermophilic model sequence, the melting temperature of subtilisin E was increased by 13°C. While wild-type subtilisin E was inactive at 90°C, the mutant retained a substantial fraction of its function, with ca. one-third of the activity that it has at 45°C.

Keywords: PROBE and Classifier; strand-turn-strand motif; subtilisin; thermostability.
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




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.