Analysis of Tat targeting function and twin-arginine signal peptide activity in Escherichia coli.

Tracy Palmer; Ben C. Berks; Frank Sargent

doi:10.1007/978-1-60327-412-8_12

Analysis of Tat targeting function and twin-arginine signal peptide activity in Escherichia coli.

Tracy Palmer, Ben C. Berks, Frank Sargent

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed)

19 Citations (Scopus)

Abstract

The Tat system is a protein export system dedicated to the transport of folded proteins across the prokaryotic cytoplasmic membrane and the thylakoid membrane of plant chloroplasts. Proteins are targeted for export by the Tat system via N-terminal signal peptides harbouring an S-R-R-x-F-L-K 'twin-arginine' motif. In this chapter qualitative and quantitative assays for native Tat substrates in the model organism Escherichia coli are described. Genetic screening methods designed to allow the rapid positive selection of Tat signal peptide activity and the first positive selection for mutations that inactivate the Tat pathway are also presented. Finally isothermal titration calorimetry (ITC) methods for measuring the affinity of twin-arginine signal peptide-chaperone interactions are discussed.

Original language	English
Title of host publication	Protein secretion
Subtitle of host publication	methods and protocols
Editors	Anastassios Economou
Publisher	Humana Press
Pages	191-216
Number of pages	26
ISBN (Electronic)	9781603274128
ISBN (Print)	9781603271677
DOIs	https://doi.org/10.1007/978-1-60327-412-8_12
Publication status	Published - 2010

Publication series

Name	Methods in Molecular Biology
Publisher	Humana Press
Volume	619
ISSN (Print)	1064-3745

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10.1007/978-1-60327-412-8_12

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abstract = "The Tat system is a protein export system dedicated to the transport of folded proteins across the prokaryotic cytoplasmic membrane and the thylakoid membrane of plant chloroplasts. Proteins are targeted for export by the Tat system via N-terminal signal peptides harbouring an S-R-R-x-F-L-K 'twin-arginine' motif. In this chapter qualitative and quantitative assays for native Tat substrates in the model organism Escherichia coli are described. Genetic screening methods designed to allow the rapid positive selection of Tat signal peptide activity and the first positive selection for mutations that inactivate the Tat pathway are also presented. Finally isothermal titration calorimetry (ITC) methods for measuring the affinity of twin-arginine signal peptide-chaperone interactions are discussed.",

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Analysis of Tat targeting function and twin-arginine signal peptide activity in Escherichia coli. / Palmer, Tracy; Berks, Ben C.; Sargent, Frank.
Protein secretion: methods and protocols. ed. / Anastassios Economou. Humana Press, 2010. p. 191-216 (Methods in Molecular Biology; Vol. 619).

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed)

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AB - The Tat system is a protein export system dedicated to the transport of folded proteins across the prokaryotic cytoplasmic membrane and the thylakoid membrane of plant chloroplasts. Proteins are targeted for export by the Tat system via N-terminal signal peptides harbouring an S-R-R-x-F-L-K 'twin-arginine' motif. In this chapter qualitative and quantitative assays for native Tat substrates in the model organism Escherichia coli are described. Genetic screening methods designed to allow the rapid positive selection of Tat signal peptide activity and the first positive selection for mutations that inactivate the Tat pathway are also presented. Finally isothermal titration calorimetry (ITC) methods for measuring the affinity of twin-arginine signal peptide-chaperone interactions are discussed.

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Analysis of Tat targeting function and twin-arginine signal peptide activity in Escherichia coli.

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