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 proceedingChapter (peer-reviewed)

    14 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 languageEnglish
    Title of host publicationProtein secretion
    Subtitle of host publicationmethods and protocols
    EditorsAnastassios Economou
    PublisherHumana Press
    Pages191-216
    Number of pages26
    ISBN (Electronic)9781603274128
    ISBN (Print)9781603271677
    DOIs
    Publication statusPublished - 2010

    Publication series

    NameMethods in Molecular Biology
    PublisherHumana Press
    Volume619
    ISSN (Print)1064-3745

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    Cite this

    Palmer, T., Berks, B. C., & Sargent, F. (2010). Analysis of Tat targeting function and twin-arginine signal peptide activity in Escherichia coli. In A. Economou (Ed.), Protein secretion: methods and protocols (pp. 191-216). (Methods in Molecular Biology; Vol. 619). Humana Press. https://doi.org/10.1007/978-1-60327-412-8_12