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Prosequence switching : An effective strategy to produce biologically active E. coli heat-stable enterotoxin STh

Title data

Weiglmeier, Philipp Richard ; Berkner, Hanna ; Seebahn, Angela ; Vogel, Nico ; Schreiber, Rainer ; Wöhrl, Birgitta M. ; Schwarzinger, Stephan ; Rösch, Paul:
Prosequence switching : An effective strategy to produce biologically active E. coli heat-stable enterotoxin STh.
In: Journal of Biomolecular Structure & Dynamics. Vol. 32 (2014) Issue 10 . - pp. 1-9.
ISSN 0739-1102
DOI: https://doi.org/10.1080/07391102.2013.825758

Abstract in another language

Enterotoxigenic Escherichia coli (ETEC) infections account for the majority of cases of acute secretory diarrhea. The causative agents are enterotoxins secreted by ETEC, among them is the heat-stable enterotoxin, STh. STh is a 19-amino acid peptide containing three disulfide bonds that stimulates fluid secretion in the bowel by binding to the receptor domain of intestinal guanylyl cyclase C (GC-C). Since GC-C agonists have pharmacologic potential for diagnosis and treatment of disorders such as constipation-predominant irritable bowel syndrome (IBS-C), chronic constipation, and colorectal carcinoma, it is crucial to develop methods for the large-scale production of STh and related peptides. Here, we present a strategy for recombinant expression of STh that relies on the use of the prosequence of human uroguanylin to support proper folding and disulfide bond formation. The chimeric protein CysCys-STh consisting of the propeptide of uroguanylin as N-terminus and the STh peptide as C-terminus was expressed in E. coli, and an efficient purification protocol was developed. Trypsin digestion of this protein released the enterotoxin which could be obtained in high purity. NMR and mass spectrometry confirmed the identity and homogeneity of the toxin, and its biological activity was confirmed by a cell-based in vivo assay. The expression scheme introduced here represents a cost-efficient and scalable way of STh production.

Further data

Item Type: Article in a journal
Refereed: Yes
Institutions of the University: Faculties
Faculties > Faculty of Biology, Chemistry and Earth Sciences
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biopolymers
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Former Professors > Chair Biopolymers - Univ.-Prof. Dr. Paul Rösch
Profile Fields > Emerging Fields > Food and Health Sciences
Profile Fields
Profile Fields > Emerging Fields
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Former Professors
Result of work at the UBT: Yes
DDC Subjects: 500 Science
500 Science > 500 Natural sciences
500 Science > 540 Chemistry
500 Science > 570 Life sciences, biology
Date Deposited: 14 Jan 2015 09:29
Last Modified: 15 May 2017 11:54
URI: https://eref.uni-bayreuth.de/id/eprint/5517