Skip Navigation


PEDS Advance Access originally published online on August 23, 2008
Protein Engineering Design and Selection 2008 21(10):631-638; doi:10.1093/protein/gzn038
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow All Versions of this Article:
21/10/631    most recent
gzn038v1
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 Jucovic, M.
Right arrow Articles by Chen, J. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Jucovic, M.
Right arrow Articles by Chen, J. S.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2008. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

From enzyme to zymogen: engineering Vip2, an ADP-ribosyltransferase from Bacillus cereus, for conditional toxicity

Milan Jucovic2, Frederick S. Walters, Gregory W. Warren, Narendra V. Palekar and Jeng S. Chen

Syngenta Biotechnology Inc., Research Triangle Park, 3054 East Cornwallis Road, NC 27709-2257, USA

2 To whom correspondence should be addressed. E-mail: milan.jucovic{at}syngenta.com

The adenosine diphosphate (ADP)-ribosyltransferase, Vip2 (vegetative insecticidal protein), from Bacillus cereus in combination with another protein from the same organism, Vip1, has insecticidal activity against western corn rootworm larvae. The Vip2 protein exerts its intracellular poisoning effect by modifying actin and preventing actin polymerization. Due to the nature of this toxin, expression of Vip2 in planta is lethal. In this work, we attempted to build an enzyme precursor (proenzyme, zymogen) that would silently reside in one biological system (e.g. plants or yeast) and be activated in the other (insect larvae). Our approach involved engineering a random propeptide library at the C-terminal end of Vip2 and selecting for malfunctional enzyme variants in yeast. A selected proenzyme (proVip2) possesses reduced enzymatic activity as compared with the wild-type Vip2 protein, but remains a potent toxin toward rootworm larvae. In addition, upon analysis of the digestive fate of the engineered enzyme precursor in rootworm larvae, we demonstrated that ‘zymogenized’ Vip2 can be proteolytically activated by rootworm digestive enzyme machinery. This report represents an example of applying a protein engineering strategy for the creation of a plant-tolerated, zymogen-like form of an otherwise toxic protein. This approach may outline a novel path to address challenges associated with utilizing toxic proteins in certain biotechnological applications.

Keywords: ADP-ribosyltransferase/conditional toxicity/proenzyme/Vip2/zymogen

Received December 12, 2007; revised May 11, 2008; accepted June 20, 2008.


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.