This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
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 Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Malone, C. L.
Right arrow Articles by Horswill, A. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Malone, C. L.
Right arrow Articles by Horswill, A. R.
Agricola
Right arrow Articles by Malone, C. L.
Right arrow Articles by Horswill, A. R.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, October 2007, p. 6036-6044, Vol. 73, No. 19
0099-2240/07/$08.00+0     doi:10.1128/AEM.00912-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein{triangledown}

Cheryl L. Malone, Blaise R. Boles, and Alexander R. Horswill*

Department of Microbiology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, Iowa 52242

Received 23 April 2007/ Accepted 1 August 2007

The Agr quorum-sensing system of Staphylococcus aureus modulates the expression of virulence factors in response to autoinducing peptides (AIPs). The peptides are seven to nine residues in length and have the C-terminal five residues constrained in a thiolactone ring. We have developed a new method to generate AIP structures using an engineered DnaB mini-intein from Synechocystis sp. strain PCC6803. In the method, an oligonucleotide encoding the AIP is ligated to the intein and the fusion protein is expressed and purified by affinity chromatography. To produce the correct AIP structure, intein splicing is interrupted, allowing the cysteine side chain to catalyze thiolactone ring formation and release AIP from the resin. The technique is simple and robust, and we have successfully produced the three main classes of AIPs using the intein system. The intein-generated AIPs possessed the correct thiolactone ring modification based on biochemical analysis, and, importantly, all the samples were bioactive against S. aureus. The AIP activity was confirmed through Agr interference and activation profiling with developed S. aureus reporter strains. The simplicity of the method, benefits of DNA encoding, and scalable nature enable the production of S. aureus AIPs for many biological applications.

* Corresponding author. Mailing address: Department of Microbiology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242. Phone: (319) 335-7783. Fax: (319) 335-8228. E-mail: alex-horswill{at}uiowa.edu

{triangledown} Published ahead of print on 10 August 2007.

Applied and Environmental Microbiology, October 2007, p. 6036-6044, Vol. 73, No. 19
0099-2240/07/$08.00+0     doi:10.1128/AEM.00912-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Thoendel, M., Horswill, A. R. (2009). Identification of Staphylococcus aureus AgrD Residues Required for Autoinducing Peptide Biosynthesis. J. Biol. Chem. 284: 21828-21838 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»